Sample records for science cell culture
-
Cell culture experiments planned for the space bioreactor
Morrison, Dennis R.; Cross, John H.
1987-01-01
Culturing of cells in a pilot-scale bioreactor remains to be done in microgravity. An approach is presented based on several studies of cell culture systems. Previous and current cell culture research in microgravity which is specifically directed towards development of a space bioprocess is described. Cell culture experiments planned for a microgravity sciences mission are described in abstract form.
-
Cell culture techniques in honey bee research
Cell culture techniques are indispensable in most if not all life science disciplines to date. Wherever cell culture models are lacking scientific development is hampered. Unfortunately this has been and still is the case in honey bee research because permanent honey bee cell lines have not yet been...
-
Cultura Obscura: Race, Power, and "Culture Talk" in the Health Sciences.
Benjamin, Ruha
2017-05-01
"The price of culture is a Lie." 1 This Article advances a critical race approach to the health sciences by examining "culture talk" as a discursive repertoire that attributes distinct beliefs, behaviors, and dispositions to ethno-racialized groups. Culture talk entails a twofold process of obfuscation - concealing the social reality of the people it describes and hiding the positionality of those who employ cultural generalizations. After tracing how culture talk circulates and reproduces racist narratives in and beyond the health sciences, I examine how cultural competency training in medical schools and diversity initiatives in stem cell research use the idiom of culture to manage and manufacture group differences. From culturing cells in the lab to enculturing people in the clinic, I apply the concept of coproduction to argue that culture talk is a precondition and product of scientific knowledge construction.
-
Forensic culture as epistemic culture: the sociology of forensic science.
Cole, Simon A
2013-03-01
This paper explores whether we can interpret the notion of 'forensic culture' as something akin to what Knorr-Cetina called an 'epistemic culture'. Can we speak of a 'forensic culture', and, if so, how is it similar to, or different from, other epistemic cultures that exist in what is conventionally called 'science'? This question has important policy implications given the National Academy Science's (NAS) recent identification of 'culture' as one of the problems at the root of what it identified as 'serious deficiencies' in U.S. forensic science and 'scientific culture' as an antidote to those problems. Finding the NAS's characterisation of 'scientific culture' overly general and naïve, this paper offers a preliminary exploration of what might be called a 'forensic culture'. Specifically, the paper explores the way in which few of the empirical findings accumulated by sociologists of science about research science seem to apply to forensic science. Instead, forensic science seems to have developed a distinct culture for which a sociological analysis will require new explanatory tools. Faithful sociological analysis of 'forensic culture' will be a necessary prerequisite for the kind of culture change prescribed by external reformist bodies like the NAS. Copyright © 2012. Published by Elsevier Ltd.
-
Is psychological science a-cultural?
Gone, Joseph P
2011-07-01
The history of psychological science, as it has intersected with ethnoracial, cultural, and other marginalized domains of group difference, is replete with disinterest, dismissal, or denigration of these diverse forms of psychological experience. This has led some to wonder whether psychological science is a-cultural, or even anti-cultural in orientation. Assessment of this provocative proposition first requires exploration of three composite questions: (1) What is culture?, (2) What is science?, and (3) What is psychological science? Based on brief consideration of these composite questions--which are remarkably complex in their own right--I argue that psychological science is not, has never been, and indeed cannot in principle be a-cultural. Instead, like all forms of knowing, psychological science emerges at particular historical moments to achieve particular goals that are motivated by particular interests. Throughout much of the history of psychological science, these goals and interests were tied to ideologically suspect agendas that contemporary psychologists are right to repudiate. The interesting question becomes whether psychology's knowledge practices can be disentangled from this earlier ideological contamination to furnish the discipline with viable methods. I propose that psychological science can in fact be so disentangled; nevertheless, the resulting methods are never adopted or deployed outside of culturally constituted interests, objectives, and motivations, thereby requiring ongoing critical engagement with the subtexts of disciplinary knowledge production. In fact, there seem to be important ways in which psychology's scientific aspirations hobble disciplinary inquiry into the human condition that has motivated multicultural psychologists to consider alternative paradigms of inquiry.
-
The evolution of chicken stem cell culture methods.
Farzaneh, M; Attari, F; Mozdziak, P E; Khoshnam, S E
2017-12-01
1. The avian embryo is an excellent model for studying embryology and the production of pharmaceutical proteins in transgenic chickens. Furthermore, chicken stem cells have the potential for proliferation and differentiation and emerged as an attractive tool for various cell-based technologies. 2. The objective of these studies is the derivation and culture of these stem cells is the production of transgenic birds for recombinant biomaterials and vaccine manufacture, drug and cytotoxicity testing, as well as to gain insight into basic science, including cell tracking. 3. Despite similarities among the established chicken stem cell lines, fundamental differences have been reported between their culture conditions and applications. Recent conventional protocols used for expansion and culture of chicken stem cells mostly depend on feeder cells, serum-containing media and static culture. 4. Utilising chicken stem cells for generation of cell-based transgenic birds and a variety of vaccines requires large-scale cell production. However, scaling up the conventional adherent chicken stem cells is challenging and labour intensive. Development of a suspension cell culture process for chicken embryonic stem cells (cESCs), chicken primordial germ cells (PGCs) and chicken induced pluripotent stem cells (ciPSCs) will be an important advance for increasing the growth kinetics of these cells. 6. This review describes various approaches and suggestions to achieve optimal cell growth for defined chicken stem cells cultures and use in future manufacturing applications.
-
Development of a microfluidic perfusion 3D cell culture system
Park, D. H.; Jeon, H. J.; Kim, M. J.; Nguyen, X. D.; Morten, K.; Go, J. S.
2018-04-01
Recently, 3-dimensional in vitro cell cultures have gained much attention in biomedical sciences because of the closer relevance between in vitro cell cultures and in vivo environments. This paper presents a microfluidic perfusion 3D cell culture system with consistent control of long-term culture conditions to mimic an in vivo microenvironment. It consists of two sudden expansion reservoirs to trap incoming air bubbles, gradient generators to provide a linear concentration, and microchannel mixers. Specifically, the air bubbles disturb a flow in the microfluidic channel resulting in the instability of the perfusion cell culture conditions. For long-term stable operation, the sudden expansion reservoir is designed to trap air bubbles by using buoyancy before they enter the culture system. The performance of the developed microfluidic perfusion 3D cell culture system was examined experimentally and compared with analytical results. Finally, it was applied to test the cytotoxicity of cells infected with Ewing’s sarcoma. Cell death was observed for different concentrations of H2O2. For future work, the developed microfluidic perfusion 3D cell culture system can be used to examine the behavior of cells treated with various drugs and concentrations for high-throughput drug screening.
-
Discovering Socio-cultural Aspects of Science Through Artworks
Güney, Burcu Gülay; Şeker, Hayati
2017-11-01
Scientific literacy is one of the primary purposes of science education which briefly focuses on using and interpreting scientific explanations, understanding science within its culture. However, science curricula emphasize science with its cognitive aspects and underestimate affective and aesthetic aspects of science. Science education needs to cover beauty of science for students to cross borders between their own culture and culture of science and to achieve the aim of scientific literacy. Relating aesthetic aspects of science with content of science and paving the way for aesthetic experiences through artworks may enrich science education. The purposes of this study are to discuss the need of integrating aesthetic aspects of science in science instruction and to propose examples and pedagogical suggestions to promote aesthetic experiences into the science education. Artworks are selected to present socio-cultural aspects of science to demonstrate the culture of science, their stories are explained, and pedagogical suggestions are proposed. Advantages and difficulties of using artworks in science instruction are discussed as a result of the study.
-
The cultural side of science communication.
Medin, Douglas L; Bang, Megan
2014-09-16
The main proposition of this paper is that science communication necessarily involves and includes cultural orientations. There is a substantial body of work showing that cultural differences in values and epistemological frameworks are paralleled with cultural differences reflected in artifacts and public representations. One dimension of cultural difference is the psychological distance between humans and the rest of nature. Another is perspective taking and attention to context and relationships. As an example of distance, most (Western) images of ecosystems do not include human beings, and European American discourse tends to position human beings as being apart from nature. Native American discourse, in contrast, tends to describe humans beings as a part of nature. We trace the correspondences between cultural properties of media, focusing on children's books, and cultural differences in biological cognition. Finally, implications for both science communication and science education are outlined.
-
Oscillating Cell Culture Bioreactor
Freed, Lisa E.; Cheng, Mingyu; Moretti, Matteo G.
2010-01-01
To better exploit the principles of gas transport and mass transport during the processes of cell seeding of 3D scaffolds and in vitro culture of 3D tissue engineered constructs, the oscillatory cell culture bioreactor provides a flow of cell suspensions and culture media directly through a porous 3D scaffold (during cell seeding) and a 3D construct (during subsequent cultivation) within a highly gas-permeable closed-loop tube. This design is simple, modular, and flexible, and its component parts are easy to assemble and operate, and are inexpensive. Chamber volume can be very low, but can be easily scaled up. This innovation is well suited to work with different biological specimens, particularly with cells having high oxygen requirements and/or shear sensitivity, and different scaffold structures and dimensions. The closed-loop changer is highly gas permeable to allow efficient gas exchange during the cell seeding/culturing process. A porous scaffold, which may be seeded with cells, is fixed by means of a scaffold holder to the chamber wall with scaffold/construct orientation with respect to the chamber determined by the geometry of the scaffold holder. A fluid, with/without biological specimens, is added to the chamber such that all, or most, of the air is displaced (i.e., with or without an enclosed air bubble). Motion is applied to the chamber within a controlled environment (e.g., oscillatory motion within a humidified 37 C incubator). Movement of the chamber induces relative motion of the scaffold/construct with respect to the fluid. In case the fluid is a cell suspension, cells will come into contact with the scaffold and eventually adhere to it. Alternatively, cells can be seeded on scaffolds by gel entrapment prior to bioreactor cultivation. Subsequently, the oscillatory cell culture bioreactor will provide efficient gas exchange (i.e., of oxygen and carbon dioxide, as required for viability of metabolically active cells) and controlled levels of fluid
-
L'esprit de sel science, culture, politique
Lévy-Leblond, Jean-Marc
1984-01-01
L'esprit de sel. Activité de recherche, la science est-elle nécessairement productrice de connaissances ? Y a-t-il de nouvelles formes et normes du savoir ? Qu'est la science pour la science ? Activité intellectuelle. la science est-elle aussi une activité culturelle ? La science moderne est-elle dans la culture. est-elle une nouvelle culture - devrait-elle l'être ? Qu'est la science pour la culture ? Activité sociale. la science est liée de façon complexe aux structures économiques et à la conjoncture politique. Quel rôle joue-t-elle, quels conditionnements subit-elle ? Qu'est la science pour la politique ? Voici un ensemble de contributions pour aiguiser ces questions cruciales et décaper les idées reçues.
-
Oers, van M.M.; Lynn, D.E.
2010-01-01
Insect cell cultures are widely used in studies on insect cell physiology, developmental biology and microbial pathology. In particular, insect cell culture is an indispensable tool for the study of insect viruses. The first continuously growing insect cell cultures were established from
-
Malina, Roger F.
2012-01-01
In recent years a number of cultural organizations have established ongoing programs of public engagement with astronomy, space science and astronautics. Many involve elements of citizen science initiatives, artists’ residencies in scientific laboratories and agencies, art and science festivals, and social network projects as well as more traditional exhibition venues. Recognizing these programs several agencies and organizations have established mechanisms for facilitating public engagement with astronomy and space science through cultural activities. The International Astronautics Federation has established an Technical Activities Committee for the Cultural Utilization of Space. Over the past year the NSF and NEA have organized disciplinary workshops to develop recommendations relating to art-science interaction and community building efforts. Rationales for encouraging public engagement via cultural projects range from theory of creativity, innovation and invention to cultural appropriation in the context of `socially robust science’ as advocated by Helga Nowotny of the European Research Council. Public engagement with science, as opposed to science education and outreach initiatives, require different approaches. Just as organizations have employed education professionals to lead education activities, so they must employ cultural professionals if they wish to develop public engagement projects via arts and culture. One outcome of the NSF and NEA workshops has been development of a rationale for converting STEM to STEAM by including the arts in STEM methodologies, particularly for K-12 where students can access science via arts and cultural contexts. Often these require new kinds of informal education approaches that exploit locative media, gaming platforms, artists projects and citizen science. Incorporating astronomy and space science content in art and cultural projects requires new skills in `cultural translation’ and `trans-mediation’ and new kinds
-
Cultural Memory Banking in Preservice Science Teacher Education
Handa, Vicente C.; Tippins, Deborah J.
2012-12-01
This study focused on the exemplification of cultural memory banking as an ethnographic tool to understand cultural practices relevant to science teaching and learning in a rural coastal village in a central island of the Philippine archipelago. Using the collaborative action ethnography as a research methodology, 10 prospective science teachers and a science teacher educator/doctoral candidate formed a research team and documented community funds of knowledge relevant to science teaching and learning through their participation in a Community Immersion course. The study employed the use of the cultural memory banking as a meditational tool to analyze, make sense of, and represent interview, focus-group discussion, and observation data, among others, for the development of culturally relevant science lessons. Originally used as an anthropological tool to preserve cultural knowledge associated with the cultivation of indigenous plant varieties, the cultural memory banking, as adapted in science education, was used, both as a data collection and analytic tool, to locate relevant science at the intersection of community life. The research team developed a cultural memory bank exemplar, "Ginamos: The Stinky Smell that Sells," to highlight the learning experiences and meaning-making process of those involved in its development. Dilemmas and insights on the development and use of cultural memory banking were discussed with respect to issues of knowledge mining and mainstreaming of indigenous/local funds of knowledge, troubling the privileged position of Western-inspired nature of science.
-
Cultural Emergence: Theorizing Culture in and from the Margins of Science Education
Wood, Nathan Brent; Erichsen, Elizabeth Anne; Anicha, Cali L.
2013-01-01
This special issue of the Journal of Research in Science Teaching seeks to explore conceptualizations of culture that address contemporary challenges in science education. Toward this end, we unite two theoretical perspectives to advance a conceptualization of culture as a complex system, emerging from iterative processes of cultural bricolage,…
-
Label-Free Raman Hyperspectral Imaging of Single Cells Cultured on Polymer Substrates.
Sinjab, Faris; Sicilia, Giovanna; Shipp, Dustin W; Marlow, Maria; Notingher, Ioan
2017-12-01
While Raman hyperspectral imaging has been widely used for label-free mapping of biomolecules in cells, these measurements require the cells to be cultured on weakly Raman scattering substrates. However, many applications in biological sciences and engineering require the cells to be cultured on polymer substrates that often generate large Raman scattering signals. Here, we discuss the theoretical limits of the signal-to-noise ratio in the Raman spectra of cells in the presence of polymer signals and how optical aberrations may affect these measurements. We show that Raman spectra of cells cultured on polymer substrates can be obtained using automatic subtraction of the polymer signals and demonstrate the capabilities of these methods in two important applications: tissue engineering and in vitro toxicology screening of drugs. Apart from their scientific and technological importance, these applications are examples of the two most common measurement configurations: (1) cells cultured on an optically thick polymer substrate measured using an immersion/dipping objective; and (2) cells cultured on a transparent polymer substrate and measured using an inverted optical microscope. In these examples, we show that Raman hyperspectral data sets with sufficient quality can be successfully acquired to map the distribution of common biomolecules in cells, such as nucleic acids, proteins, and lipids, as well as detecting the early stages of apoptosis. We also discuss strategies for further improvements that could expand the application of Raman hyperspectral imaging on polymer substrates even further in biomedical sciences and engineering.
-
Home Culture, Science, School and Science Learning: Is Reconciliation Possible?
Tan, Aik-Ling
2011-01-01
In response to Meyer and Crawford's article on how nature of science and authentic science inquiry strategies can be used to support the learning of science for underrepresented students, I explore the possibly of reconciliation between the cultures of school, science, school science as well as home. Such reconciliation is only possible when…
-
Political science factor in information culture
Baranov G.
2017-01-01
The value of political science in information culture of society reveals; the main indicators of the public status of political science are investigated; the main functions of political science in the activity of actors of society are characterised.
-
21st Century Cell Culture for 21st Century Toxicology.
Pamies, David; Hartung, Thomas
2017-01-17
There is no good science in bad models. Cell culture is especially prone to artifacts. A number of novel cell culture technologies have become more broadly available in the 21st century, which allow overcoming limitations of traditional culture and are more physiologically relevant. These include the use of stem-cell derived human cells, cocultures of different cell types, scaffolds and extracellular matrices, perfusion platforms (such as microfluidics), 3D culture, organ-on-chip technologies, tissue architecture, and organ functionality. The physiological relevance of such models is further enhanced by the measurement of biomarkers (e.g., key events of pathways), organ specific functionality, and more comprehensive assessment cell responses by high-content methods. These approaches are still rarely combined to create microphysiological systems. The complexity of the combination of these technologies can generate results closer to the in vivo situation but increases the number of parameters to control, bringing some new challenges. In fact, we do not argue that all cell culture needs to be that sophisticated. The efforts taken are determined by the purpose of our experiments and tests. If only a very specific molecular target to cell response is of interest, a very simple model, which reflects this, might be much more suited to allow standardization and high-throughput. However, the less defined the end point of interest and cellular response are, the better we should approximate organ- or tissue-like culture conditions to make physiological responses more probable. Besides these technologic advances, important progress in the quality assurance and reporting on cell cultures as well as the validation of cellular test systems brings the utility of cell cultures to a new level. The advancement and broader implementation of Good Cell Culture Practice (GCCP) is key here. In toxicology, this is a major prerequisite for meaningful and reliable results, ultimately
-
Teaching Science from Cultural Points of Intersection
Grimberg, Bruna Irene; Gummer, Edith
2013-01-01
This study focuses on a professional development program for science teachers near or on American Indian reservations in Montana. This program was framed by culturally relevant pedagogy premises and was characterized by instructional strategies and content foci resulting from the intersection between three cultures: tribal, science teaching, and…
-
Stampfer, Martha R; Garbe, James C
2015-02-24
Cell culture media formulations for culturing human epithelial cells are herein described. Also described are methods of increasing population doublings in a cell culture of finite life span human epithelial cells and prolonging the life span of human cell cultures. Using the cell culture media disclosed alone and in combination with addition to the cell culture of a compound associated with anti-stress activity achieves extended growth of pre-stasis cells and increased population doublings and life span in human epithelial cell cultures.
-
When technology, science and culture meet: insights from ancient Chinese technology
Lee, Yeung Chung
2017-10-01
This paper draws together two important agendas in science education. The first is making science education more inclusive such that students from non-Western or indigenous cultures can benefit from culturally relevant curricula. The second is integrating technology into the curriculum under the umbrella of Science-Technology-Society (STS) education to embrace the social aspects of science, with technology serving as a bridge. The advancement of the first agenda is hindered by the pursuance by both Western and non-Western societies of narrow cultural and practical goals without considering the development of science and technology from a cross-cultural perspective. The second agenda is limited by the misconception that technology is applied science, leading to the exclusion from STS discussions of pre-science or indigenous technologies developed by non-Western cultures. Through selected case studies of the evolution of Chinese traditional technologies and their interaction with science, this paper offers a perspective from the Far East, and argues for situating culturally responsive science education in broader historical and cross-cultural contexts to acknowledge the multi-cultural contributions to science and technology. A form of cross-cultural STS education is advanced, encompassing the cultural basis of technological developments, technology diffusion, interactions of traditional technology with science, and the potential development of traditional or indigenous technologies. This approach provides a bridge between the existing universal science education paradigm promoted in the West and the different forms of multi-cultural education advocated by indigenous science educators. To translate theory into practice, a conceptual framework is proposed in which the essential transdisciplinary knowledge base, curricular goals, and pedagogical approaches are embedded.
-
Toward a Psychological Science for a Cultural Species.
Heine, Steven J; Norenzayan, Ara
2006-09-01
Humans are a cultural species, and the study of human psychology benefits from attention to cultural influences. Cultural psychology's contributions to psychological science can largely be divided according to the two different stages of scientific inquiry. Stage 1 research seeks cultural differences and establishes the boundaries of psychological phenomena. Stage 2 research seeks underlying mechanisms of those cultural differences. The literatures regarding these two distinct stages are reviewed, and various methods for conducting Stage 2 research are discussed. The implications of culture-blind and multicultural psychologies for society and intergroup relations are also discussed. © 2006 Association for Psychological Science.
-
Lloyd Roberts memorial lecture. Science, culture and wealth.
Noble, D
1. This lecture defends the view that science, culture and wealth are linked and that, in the long run, the only way to maintain the spirit of excited intellectual enquiry leading to novel exploitable ideas is to attract the young by creating and maintaining a culture in which they respond to the intellectual challenge. 2. The pursuit of science is not independent of the culture in which it develops, nor is it a neutral activity. The objectivity of science neither requires nor entails its neutrality. 3. A comparison between American and British attitudes reveals a major cultural difference that leads otherwise similar political authorities to totally opposed views on the role of public funding of science. 4. The roots of this difference run deep in our culture and have a long history, stretching back at least to the early 19th century and Babbage's Decline of Science campaign. This seems to be a feature of the culture of the governing classes in Britain, at least in modern Britain. The general public perception (revealed by a recent opinion poll) is that more than 80% think that our national prosperity depends on science and technology and that it is important for Britain to be a leading nation in science. 5. The immediate cause of the present political malaise with regard to science funding is the perceived lack of correlation between science expenditure and industrial success in the 1960s. In fact, though, at the micro-economic level, there is a strong correlation between research investment and industrial competitiveness. Those industries that have invested have also succeeded. The general problem lies with a failure of major parts of industry to invest in research rather than in any major weakness or lack of exploitability in British science. 6. It will not solve that problem (which to compare with our main competitors requires an increased civil Research and Development expenditure of 3 pounds billion/annum by British government and industry combined) to try
-
Preparing teachers for ambitious and culturally responsive science teaching
Seiler, Gale
2013-03-01
Communities, schools and classrooms across North America are becoming more ethnically, racially, and linguistically diverse, particularly in urban areas. Against this backdrop, underrepresentation of certain groups in science continues. Much attention has been devoted to multicultural education and the preparation of teachers for student diversity. In science education, much research has focused on classrooms as cultural spaces and the need for teachers to value and build upon students' everyday science knowledge and ways of sense-making. However it remains unclear how best to prepare science teachers for this kind of culturally responsive teaching. In attempting to envision how to prepare science teachers with cross-cultural competency, we can draw from a parallel line of research on preparing teachers for ambitious science instruction. In ambitious science instruction, students solve authentic problems and generate evidence and models to develop explanations of scientific phenomenon, an approach that necessitates great attention to students' thinking and sense-making, thus making it applicable to cultural relevance aims. In addition, this line of research on teacher preparation has developed specific tools and engages teachers in cycles of reflection and rehearsal as they develop instructional skills. While not addressing cross-cultural teaching specifically, this research provides insights into specific ways through which to prepare teachers for culturally responsive practices. In my presentation, I will report on efforts to join these two areas of research, that is, to combine ideas about multicultural science teacher preparation with what has been learned about how to develop ambitious science instruction. This research suggests a new model for urban science teacher preparation--one that focuses on developing specific teaching practices that elicit and build on student thinking, and doing so through cycles of individual and collective planning, rehearsal
-
Indian Science Culture Needs a Paradigm Shift.
Sharma, Om P
2017-06-01
There is a general impression in the scientific community in our country that the way science is taught, leant and the work culture of research and management of academic and research institutions is not conducive to cutting edge research, innovation and making world leaders. Mentoring continues to be poor with some exceptions. Very often, senior scientists with long innings in science teaching and research express anguish at the status quo in spite of a number of policy documents and recommendations for change. Indian science culture (teaching, research as well as administration) is a matter of prime concern and the issues cannot be pushed under the carpet if we desire a qualitative change. Most of the institutions of higher learning churn out graduates and post graduates who are largely unemployable. There are concerns on the number of Ph.Ds and not on the quality of Ph.D. One major consequence of the weak post graduates and Ph.Ds is the non-availability of competent faculty. Weakness and lack of interest in science learning starts from school. Learning continues to be by rote which is the prime reason for our low global rank in science and mathematics competence. Teaching and research apart there are umpteen other issues in over all culture of institutions and universities engaged in science teaching and research. Few oases of excellence are exceptions in the vast pool of mediocrity. Some points which need prime attention are: adoption of a tenure track system on the pattern of US institutions; feedback on and evaluation of teaching and mentoring; bottom up approach for candid feedback on issues which require long term solutions for efficiency and sound deliverables, cultivating the culture of working in front line areas, full transparency in working and an all out exit from culture of feudalism. This transformation needs commitment on the part of the politicians who man the respective departments of science education, research and human resource development. I
-
Applied linguistics - a science of culture?
Directory of Open Access Journals (Sweden)
Benke, Gertraud
2003-01-01
Full Text Available In this article, the status of applied linguistics as discipline is questioned and problems of establishing it - and other newly formed scientific enterprises like cultural science - as disciplines are discussed. This discussion is contextualized using the author's own experience as applied linguist working in (the institutional structure of Austria. Secondly, applied linguistics is presented as complementing cultural science, with both exploring at times the same phenomena albeit under different perspectives and focussing on different levels of experience. Two examples of research involving such a joint interest with different foci are discussed.
-
Supporting pre-service science teachers in developing culturally relevant pedagogy
Krajeski, Stephen
This study employed a case study methodology to investigate a near-authentic intervention program designed to support the development of culturally relevant pedagogy and its impact on pre-service science teachers' notions of culturally relevant pedagogy. The unit of analysis for this study was the discourse of pre-service science teachers enrolled in a second semester science methods course, which was the site of the intervention program. Data for this study was collected from videos of classroom observations, audio recordings of personal interviews, and artifacts created by the pre-service science teachers during the class. To determine how effective science teacher certification programs are at supporting the development of culturally relevant pedagogy without an immersion aspect, two research questions were investigated: 1) How do pre-service science teachers view and design pedagogy while participating in an intervention designed to support the development of culturally relevant pedagogy? 2) How do pre-service science teachers view the importance of culturally relevant pedagogy for supporting student learning? How do their practices in the field change these initial views?
-
Cultural, Social and Political Perspectives in Science Education
DEFF Research Database (Denmark)
education research to question whether conventional research approaches, foci and theoretical approaches are sufficient in a world of science education that is neither politically neutral, nor free of cultural values. Attention is not only on the individual learner but on the cultural, social and political......This book presents a collection of critical thinking that concern cultural, social and political issues for science education in the Nordic countries. The chapter authors describe specific scenarios to challenge persisting views, interrogate frameworks and trouble contemporary approaches...... to researching teaching and learning in science. Taking a point of departure in empirical examples from the Nordic countries the collection of work is taking a critical sideways glance at the Nordic education principles. Critical examinations target specifically those who are researching in the fields of science...
-
Cultural Memory Banking in Preservice Science Teacher Education
Handa, Vicente C.; Tippins, Deborah J.
2012-01-01
This study focused on the exemplification of cultural memory banking as an ethnographic tool to understand cultural practices relevant to science teaching and learning in a rural coastal village in a central island of the Philippine archipelago. Using the collaborative action ethnography as a research methodology, 10 prospective science teachers…
-
Baert, Yoni; Braye, Aude; Struijk, Robin B.; van Pelt, Ans M. M.; Goossens, Ellen
2015-01-01
To assess whether testicular cell dynamics are altered during long-term culture after testicular tissue cryopreservation. Experimental basic science study. Reproductive biology laboratory. Testicular tissue with normal spermatogenesis was obtained from six donors. None. Detection and comparison of
-
Investigating Your School's Science Teaching and Learning Culture
Sato, Mistilina; Bartiromo, Margo; Elko, Susan
2016-01-01
The authors report on their work with the Academy for Leadership in Science Instruction, a program targeted to help science teachers promote a science teaching and learning culture in their own schools.
-
Perfusion based cell culture chips
DEFF Research Database (Denmark)
Heiskanen, Arto; Emnéus, Jenny; Dufva, Martin
2010-01-01
Performing cell culture in miniaturized perfusion chambers gives possibilities to experiment with cells under near in vivo like conditions. In contrast to traditional batch cultures, miniaturized perfusion systems provide precise control of medium composition, long term unattended cultures...... and tissue like structuring of the cultures. However, as this chapter illustrates, many issues remain to be identified regarding perfusion cell culture such as design, material choice and how to use these systems before they will be widespread amongst biomedical researchers....
-
Neural stem cell isolation and culture from C57BL/6 mice
Directory of Open Access Journals (Sweden)
S Koirala
2015-07-01
Full Text Available INTRODUCTION A widely used in vitro culture, the neurosphere assay (NSA has provided a means to retrospectively identify neural progenitor cells as well as to determine both their selfrenewal capacity. Objective of study was to isolate and compare growth of the embryonic neuronal stem cell and adult neuronal stem cells in presence of Epidermal Growth Factor (EGF and Fibroblastic Growth Factor (FGF2. MATERIALS AND METHODS Embryonic neuronal stem cell were collected from cortical plate of dorsal telencephalon of fifteen C57BL/6 transgenic mice using stereoscopic microscope on 11th gestational day (GD. Adult mammalian neuronal stem cells taken from subventricular zone (SVZ of the lateral ventricles and subgranular layer of the dentate gyrus of the hippocampus were cultured. The growth for the neurosphere was then observed in interval of 24 and 72 hours. RESULT The adult stem cell culture showed few intact cells with high amount of debris and 9% heterogeneous sphere after 24 hours while only 20 % was observed at the end of 72 hours. Higher proliferation rate was observed in embryonic neurospheres than the adult stem cell culture. CONCLUSION Presence of EGF and basic FGF2 is essential for culture of neurospheres.DOI: http://dx.doi.org/10.3126/jcmsn.v10i2.12946 Journal of College of Medical Sciences-Nepal, 2014, Vol.10(2; 1-3
-
Dye, Frank J.
1985-01-01
Outlines steps to generate cell samples for observation and experimentation. The procedures (which use ordinary laboratory equipment) will establish a short-term primary culture of normal mammalian cells. Information on culture vessels and cell division and a list of questions to generate student interest and involvement in the topics are…
-
Strategic management cultures: historical connections with science
Abreu Pederzini, G.
2016-01-01
Purpose: The implicit and indirect influence of classical science on strategic management has been of utmost importance in the development of the discipline. Classical science has underpinned the main and even contrasting strategic management cultures. Classical science has undoubtedly allowed strategic management to thrive. Nevertheless, important limitations, roadblocks and challenges have also been produced. This paper aims to explore the influence of classical science on the main positivi...
-
Science fiction as a culture of global innovation
Directory of Open Access Journals (Sweden)
Thomas MICHAUD
2010-01-01
Full Text Available Science fiction participates to the creation of a global culture of innovation. It is diffused in most of the developed countries to promote technical innovation and has motivated a lot of actors of capitalism to imitate the utopian technologies represented in these very popular movies and novels. The stake of this article is to define the strategic habitus in a cultural environment constituted of multiple centers of Research and Development (R&D organized in network. The management of science fiction is necessary to optimize innovation at a global level. After the step of the ideological filtering of science fiction, the construction of discursive philters permits to manage productive systems with common and normalized cultural considerations. The approaches of sensemaking, storytelling and “strategy as discourse” are used at the theoretical level.
-
Influence of culture and language sensitive physics on science attitude enhancement
Morales, Marie Paz E.
2015-12-01
The study critically explored how culture and language sensitive curriculum materials in physics improve Pangasinan learners' attitude towards science. Their cultural dimensions, epistemological beliefs, and views on integration of culture and language in the teaching and learning process determined their cultural preference or profile. Design and development of culture and language sensitive curriculum materials in physics were heavily influenced by these learners' cultural preference or profile. Pilot-study using interviews and focus group discussions with natives of Pangasinan and document analysis were conducted to identify the culture, practices, and traditions integrated in the lesson development. Comparison of experimental participants' pretest and posttest results on science attitude measure showed significant statistical difference. Appraisal of science attitude enhancement favored the experimental group over the control group. Qualitative data deduced from post implementation interviews, focus group discussions, and journal log entries showed the same trend in favor of the experimental participants. The study revealed that culture and language integration in the teaching and learning process of physics concepts enabled students to develop positive attitude to science, their culture, and native language.
-
Religiosity, Culture, and Science Communication
O'Malley, R. C.; Kahan, D.
2017-12-01
It is well established that cultural commitments influence receptivity to scientific information on risks and related policy-relevant facts. Religiosity is one proxy for such commitments. My presentation will present data from numerous studies (observational and experimental, lab and field) that address how religiosity as a form of cultural affinity shapes engagement with the best available evidence on human-caused climate change. The central conclusion of this research is that a skeptical position on climate change, much like a skeptical position on human evolution, operates as a tacit badge of membership in and loyalty to groups bound together by religious affiliations. Overcoming the distorting impact that this dynamic has on climate-science communication requires engaging members of religious groups not as members of those groups per se but as citizens with a practical stake in addressing the risks that climate change poses to them and their neighbors. Once enlisted into discussion and practical action on these grounds, however, religious individuals can be expected to share their positive experiences and outlooks with other members of their religious communities, thereby demonstrating to them that engaging with this form of science does not conflict with their cultural identities.
-
DEFF Research Database (Denmark)
Stangegaard, Michael; Petronis, Sarunas; Jørgensen, Anders Michael
2006-01-01
culture chip compared to cell culture flasks. The cell culture chip could without further modification support cell growth of two other cell lines. Light coming from the microscope lamp during optical recordings of the cells was the only external factor identified, that could have a negative effect...... on cell survival. Low grade light exposure was however compatible with optical recordings as well as cell viability. These results strongly indicate that a cell culture chip could be constructed that allowed for on-line optical recording of cellular events without affecting the cell culturing condition...
-
DEFF Research Database (Denmark)
Stangegaard, Michael; Petronis, Sarunas; Jørgensen, A M
2006-01-01
culture chip compared to cell culture flasks. The cell culture chip could without further modification support cell growth of two other cell lines. Light coming from the microscope lamp during optical recordings of the cells was the only external factor identified, that could have a negative effect...... on cell survival. Low grade light exposure was however compatible with optical recordings as well as cell viability. These results strongly indicate that a cell culture chip could be constructed that allowed for on-line optical recording of cellular events without affecting the cell culturing condition...
-
Directory of Open Access Journals (Sweden)
Ali Rabbani
2014-03-01
Full Text Available The main characteristic (feature of the sociology of knowledge and science is its emphasis on the culture and cultural analysis within the scientific and technological research. This study concerns with the study of two research fields in which new sociologists of science and technology have presented their cultural analysis. These two fields include: sociology of scientific knowledge and cultural studies of science.Sociology of scientific knowledge is the first school of thought which makes the content of scientific knowledge inclined to and compliant with the cultural and sociological analysis. In SSK, the main presupposition is that “the scientific knowledge is totally arbitrary.” Accordingly, the design and evaluation of scientific theories and claims are the consequence of social interests and cultural inclinations (trends, in a way that the scientific theories become a tool for the justification, legitimating, encouragement and contentment.At the early 1990s, with the rise of crisis (chaos within the explanations of sociology of scientific knowledge and a flood of criticism against it, the whole subjectivity of the field came to a standstill (reached an impasse and the initiatives in scientific research were replaced by different theoretical orientations like cultural studies. In contrast to the sociology of scientific knowledge, the cultural studies of science concerns with the rejection of “explanation” and, instead, focuses on the “meaning” and “understanding”. In other words, it has come back to an old dispute between explanatory and hermeneutic approaches and those which pursue the regulative (legalistic comprehensiveness along the more positivistic lines.This emerging field emphasizes the issue that the uncertainty, instability, ambiguity (vagueness and difference must be given a more important role in sciences. Cultural studies of science gave rise to a change from the sociology of scientific knowledge to a new
-
Pamies, David; Bal-Price, Anna; Chesné, Christophe; Coecke, Sandra; Dinnyes, Andras; Eskes, Chantra; Grillari, Regina; Gstraunthaler, Gerhard; Hartung, Thomas; Jennings, Paul; Leist, Marcel; Martin, Ulrich; Passier, Robert; Schwamborn, Jens C; Stacey, Glyn N; Ellinger-Ziegelbauer, Heidrun; Daneshian, Mardas
2018-04-13
A major reason for the current reproducibility crisis in the life sciences is the poor implementation of quality control measures and reporting standards. Improvement is needed, especially regarding increasingly complex in vitro methods. Good Cell Culture Practice (GCCP) was an effort from 1996 to 2005 to develop such minimum quality standards also applicable in academia. This paper summarizes recent key developments in in vitro cell culture and addresses the issues resulting for GCCP, e.g. the development of induced pluripotent stem cells (iPSCs) and gene-edited cells. It further deals with human stem-cell-derived models and bioengineering of organo-typic cell cultures, including organoids, organ-on-chip and human-on-chip approaches. Commercial vendors and cell banks have made human primary cells more widely available over the last decade, increasing their use, but also requiring specific guidance as to GCCP. The characterization of cell culture systems including high-content imaging and high-throughput measurement technologies increasingly combined with more complex cell and tissue cultures represent a further challenge for GCCP. The increasing use of gene editing techniques to generate and modify in vitro culture models also requires discussion of its impact on GCCP. International (often varying) legislations and market forces originating from the commercialization of cell and tissue products and technologies are further impacting on the need for the use of GCCP. This report summarizes the recommendations of the second of two workshops, held in Germany in December 2015, aiming map the challenge and organize the process or developing a revised GCCP 2.0.
-
Monteiro, Anna Karina
Research acknowledges that if students are to be successful science, they must learn to navigate and cross cultural borders that exist between their own cultures and the subculture of science. This dissertation utilized a mixed methods approach to explore how inservice science teachers working in urban schools construct their ideas of and apply the concepts about the culture of science and cultural border crossing as relevant to the teaching and learning of science. The study used the lenses of cultural capital, social constructivism, and cultural congruency in the design and analysis of each of the three phases of data collection. Phase I identified the perspectives of six inservice science teachers on science culture, cultural border crossing, and which border crossing methods, if any, they used during science teaching. Phase II took a dialectical approach as the teachers read about science culture and cultural border crossing during three informal professional learning community meetings. This phase explored how teachers constructed their understanding of cultural border crossing and how the concept applied to the teaching and learning of science. Phase III evaluated how teachers' perspectives changed from Phase I. In addition, classroom observations were used to determine whether teachers' practices in their science classrooms changed from Phase I to Phase III. All three phases collected data through qualitative (i.e., interviews, classroom observations, and surveys) and quantitative (Likert items) means. The findings indicated that teachers found great value in learning about the culture of science and cultural border crossing as it pertained to their teaching methods. This was not only evidenced by their interviews and surveys, but also in the methods they used in their classrooms. Final conclusions included how the use of student capital resources (prior experiences, understandings and knowledge, ideas an interests, and personal beliefs), if supported by
-
SCIENCE WHERE CULTURE MATTERS: A NEO-CLASSICAL ...
Indian Academy of Sciences (India)
Table of contents. SCIENCE WHERE CULTURE MATTERS: A NEO-CLASSICAL APPROACH TO EXPLORE UNTAPPED BACTERIAL DIVERSITY · UNDER GRADUATE RESEARCH An alternative model of doing science · THE EXPANSE OF LIFE · HOW MANY SP. OF BACTERIA IN 1 g SOIL? TORSVIK ET AL 1990.
-
Suspended Cell Culture ANalysis (SCAN) Tool to Enhance ISS On-Orbit Capabilities, Phase I
National Aeronautics and Space Administration — Aurora Flight Sciences and partner, Draper Laboratory, propose to develop an on-orbit immuno-based label-free Suspension Cell Culture ANalysis tool, SCAN tool, which...
-
A Conceptual Culture Model for Design Science Research
Directory of Open Access Journals (Sweden)
Thomas Richter
2016-03-01
Full Text Available The aim of design science research (DSR in information systems is the user-centred creation of IT-artifacts with regard to specific social environments. For culture research in the field, which is necessary for a proper localization of IT-artifacts, models and research approaches from social sciences usually are adopted. Descriptive dimension-based culture models most commonly are applied for this purpose, which assume culture being a national phenomenon and tend to reduce it to basic values. Such models are useful for investigations in behavioural culture research because it aims to isolate, describe and explain culture-specific attitudes and characteristics within a selected society. In contrast, with the necessity to deduce concrete decisions for artifact-design, research results from DSR need to go beyond this aim. As hypothesis, this contribution generally questions the applicability of such generic culture dimensions’ models for DSR and focuses on their theoretical foundation, which goes back to Hofstede’s conceptual Onion Model of Culture. The herein applied literature-based analysis confirms the hypothesis. Consequently, an alternative conceptual culture model is being introduced and discussed as theoretical foundation for culture research in DSR.
-
Cell Culturing of Cytoskeleton
2004-01-01
Biomedical research offers hope for a variety of medical problems, from diabetes to the replacement of damaged bone and tissues. Bioreactors, which are used to grow cells and tissue cultures, play a major role in such research and production efforts. Cell culturing, such as this bone cell culture, is an important part of biomedical research. The BioDyn payload includes a tissue engineering investigation. The commercial affiliate, Millenium Biologix, Inc., has been conducting bone implant experiments to better understand how synthetic bone can be used to treat bone-related illnesses and bone damaged in accidents. On STS-95, the BioDyn payload will include a bone cell culture aimed to help develop this commercial synthetic bone product. Millenium Biologix, Inc., is exploring the potential for making human bone implantable materials by seeding its proprietary artificial scaffold material with human bone cells. The product of this tissue engineering experiment using the Bioprocessing Modules (BPMs) on STS-95 is space-grown bone implants, which could have potential for dental implants, long bone grafts, and coating for orthopedic implants such as hip replacements.
-
2010-01-01
... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Cell cultures. 101.6 Section 101.6..., SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS DEFINITIONS § 101.6 Cell cultures. When used in conjunction with or in reference to cell cultures, which may be referred to as tissue cultures...
-
Science knowledge and cognitive strategy use among culturally and linguistically diverse students
Lee, Okhee; Fradd, Sandra H.; Sutman, Frank X.
Science performance is determined, to a large extent, by what students already know about science (i.e., science knowledge) and what techniques or methods students use in performing science tasks (i.e., cognitive strategies). This study describes and compares science knowledge, science vocabulary, and cognitive strategy use among four diverse groups of elementary students: (a) monolingual English Caucasian, (b) African-American, (c) bilingual Spanish, and (d) bilingual Haitian Creole. To facilitate science performance in culturally and linguistically congruent settings, the study included student dyads and teachers of the same language, culture, and gender. Science performance was observed using three science tasks: weather phenomena, simple machines, and buoyancy. Data analysis involved a range of qualitative methods focusing on major themes and patterns, and quantitative methods using coding systems to summarize frequencies and total scores. The findings reveal distinct patterns of science knowledge, science vocabulary, and cognitive strategy use among the four language and culture groups. The findings also indicate relationships among science knowledge, science vocabulary, and cognitive strategy use. These findings raise important issues about science instruction for culturally and linguistically diverse groups of students.Received: 3 January 1995;
-
How Our Culture Keeps out of Science
Wood, Peter
2008-01-01
In this article, the author points out that the cultural bias against serious study of science and technology is rarely recognized as a reason for American students' poor performance. Students respond more profoundly to cultural imperatives than to market forces. In the United States, students are insulated from the commercial market's demand for…
-
When Technology, Science and Culture Meet: Insights from Ancient Chinese Technology
Lee, Yeung Chung
2018-01-01
This paper draws together two important agendas in science education. The first is making science education more inclusive such that students from non-Western or indigenous cultures can benefit from culturally relevant curricula. The second is integrating technology into the curriculum under the umbrella of Science-Technology-Society (STS)…
-
Course Syllabus--Culture, Science and Technology.
Coleman, Sam
1988-01-01
Presents a course syllabus and requirements for an anthropology course on the cross-cultural analysis of the relationships between technology, science, and social organization. Provides daily topics, suggested text readings, and reference articles. (MVL)
-
Cultural, Social and Political Perspectives in Science Education
DEFF Research Database (Denmark)
This book presents a collection of critical thinking that concern cultural, social and political issues for science education in the Nordic countries. The chapter authors describe specific scenarios to challenge persisting views, interrogate frameworks and trouble contemporary approaches to resea......This book presents a collection of critical thinking that concern cultural, social and political issues for science education in the Nordic countries. The chapter authors describe specific scenarios to challenge persisting views, interrogate frameworks and trouble contemporary approaches...... to researching teaching and learning in science. Taking a point of departure in empirical examples from the Nordic countries the collection of work is taking a critical sideways glance at the Nordic education principles. Critical examinations target specifically those who are researching in the fields of science...... conditions and contexts in science education. The different chapters review debates and research in teacher education, school teaching and learning including when external stakeholders are involved. Even though the chapters are contextualized in Nordic settings there will be similarities and parallels...
-
Marinez, Diana I.; Ortiz de Montellano, Bernardo R.
There are many ways in which science can be made culturally relevant: archeoastronomy, mathematics, geology, ethnobotany, chemistry, and art can all be taught from a perspective celebrating the accomplishments of Mexican American and American Indian science and encouraging exploration. A culturally relevant curriculum provides teachers with…
-
New Metaphors about Culture: Implications for Research in Science Teacher Preparation
Seiler, Gale
2013-01-01
Culture has been commonly used in science education research, in particular to examine issues of equity for students from low-income, racial, and ethnic minority communities. It has provided a lens with which to appreciate science classrooms as cultural places and to recognize the importance of students' cultural ways of being as resources for…
-
Cultural politics: Linguistic identity and its role as gatekeeper in the science classroom
Hilton-Brown, Bryan Anthony
This dissertation investigated how participation in the cultural practices of science classrooms creates intrapersonal conflict for ethnic minority students. Grounded in research perspectives of cultural anthropology, sociocultural studies of science education, and critical pedagogy, this study examined the cultural tensions encountered by minority students as they assimilate into the culture of the science classroom. Classroom interaction was viewed from the perspective of instructional congruence---the active incorporation of students' culture into science pedagogy. Ogbu's notion of "oppositional identity", Fordham's "fictive kinship", Bahktin's "antidialogics", and Freire's "critical consciousness" were brought together to examine how members of marginalized cultures develop non-normative behaviors as a means of cultural resistance. Choice of genre for public discourse was seen as a political act, representing students' own cultural affiliations. Conducted in a diverse Southern Californian high school with an annual population of over 3,900 students, this study merged ethnographic research, action research, and sociolinguistic discourse analysis. Post hoc analysis of videotaped classroom activities, focus group interviews, and samples of student work revealed students' discursive behavior to shift as a product of the context of their discursive exchanges. In whole class discussions students explained their understanding of complex phenomena to classmates, while in small group discussions they favored brief exchanges of group data. Four domains of discursive identities were identified: Opposition Status, Maintenance Status, Incorporation Status, and Proficiency Status. Students demonstrating Opposition Status avoided use of science discourse. Those students who demonstrated Maintenance Status were committed to maintaining their own discursive behavior. Incorporation Status students were characterized by an active attempt to incorporate science discourse into
-
Microfluidic cell culture systems for drug research.
Wu, Min-Hsien; Huang, Song-Bin; Lee, Gwo-Bin
2010-04-21
In pharmaceutical research, an adequate cell-based assay scheme to efficiently screen and to validate potential drug candidates in the initial stage of drug discovery is crucial. In order to better predict the clinical response to drug compounds, a cell culture model that is faithful to in vivo behavior is required. With the recent advances in microfluidic technology, the utilization of a microfluidic-based cell culture has several advantages, making it a promising alternative to the conventional cell culture methods. This review starts with a comprehensive discussion on the general process for drug discovery and development, the role of cell culture in drug research, and the characteristics of the cell culture formats commonly used in current microfluidic-based, cell-culture practices. Due to the significant differences in several physical phenomena between microscale and macroscale devices, microfluidic technology provides unique functionality, which is not previously possible by using traditional techniques. In a subsequent section, the niches for using microfluidic-based cell culture systems for drug research are discussed. Moreover, some critical issues such as cell immobilization, medium pumping or gradient generation in microfluidic-based, cell-culture systems are also reviewed. Finally, some practical applications of microfluidic-based, cell-culture systems in drug research particularly those pertaining to drug toxicity testing and those with a high-throughput capability are highlighted.
-
Sutter, Leo; Kolbman, Dan; Wu, Mingming; Ma, Minglin; Das, Moumita
The biophysics of cell co-cultures, i.e. binary systems of cell populations, is of great interest in many biological processes including formation of embryos, and tumor progression. During these processes, different types of cells with different physical properties are mixed with each other, with important consequences for cell-cell interaction, aggregation, and migration. The role of the differences in their physical properties in their collective behavior remains poorly understood. Furthermore, until recently most theoretical studies of collective cell migration have focused on two dimensional systems. Under physiological conditions, however, cells often have to navigate three dimensional and confined micro-environments. We study a confined, three-dimensional binary system of interacting, active, and deformable particles with different physical properties such as deformability, motility, adhesion, and division rates using Langevin Dynamics simulations. Our findings may provide insights into how the differences in and interplay between cell mechanical properties, division, and motility influence emergent collective behavior such as cell aggregation and segregation experimentally observed in co-cultures of breast cancer cells and healthy breast epithelial cells. This work was partially supported by a Cottrell College Science Award.
-
Hatton, J. P.; Lewis, M. L.; Roquefeuil, S. B.; Chaput, D.; Cazenave, J. P.; Schmitt, D. A.
1998-01-01
The results of experiments performed in recent years on board facilities such as the Space Shuttle/Spacelab have demonstrated that many cell systems, ranging from simple bacteria to mammalian cells, are sensitive to the microgravity environment, suggesting gravity affects fundamental cellular processes. However, performing well-controlled experiments aboard spacecraft offers unique challenges to the cell biologist. Although systems such as the European 'Biorack' provide generic experiment facilities including an incubator, on-board 1-g reference centrifuge, and contained area for manipulations, the experimenter must still establish a system for performing cell culture experiments that is compatible with the constraints of spaceflight. Two different cell culture kits developed by the French Space Agency, CNES, were recently used to perform a series of experiments during four flights of the 'Biorack' facility aboard the Space Shuttle. The first unit, Generic Cell Activation Kit 1 (GCAK-1), contains six separate culture units per cassette, each consisting of a culture chamber, activator chamber, filtration system (permitting separation of cells from supernatant in-flight), injection port, and supernatant collection chamber. The second unit (GCAK-2) also contains six separate culture units, including a culture, activator, and fixation chambers. Both hardware units permit relatively complex cell culture manipulations without extensive use of spacecraft resources (crew time, volume, mass, power), or the need for excessive safety measures. Possible operations include stimulation of cultures with activators, separation of cells from supernatant, fixation/lysis, manipulation of radiolabelled reagents, and medium exchange. Investigations performed aboard the Space Shuttle in six different experiments used Jurkat, purified T-cells or U937 cells, the results of which are reported separately. We report here the behaviour of Jurkat and U937 cells in the GCAK hardware in ground
-
A cogenerative inquiry using postcolonial theory to envisage culturally inclusive science education
Adams, Jennifer; Luitel, Bal Chandra; Afonso, Emilia; Taylor, Peter Charles
2008-12-01
This forum constitutes a cogenerative inquiry using postcolonial theory drawn from the review paper by Zembylas and Avraamidou. Three teacher educators from African, Asian and Caribbean countries reflect on problems confronting their professional practices and consider the prospects of creating culturally inclusive science education. We learn that in Mozambique, Nepal and the Caribbean scientism patrols the borders of science education serving to exclude local epistemological beliefs and discourses and negating culturally contextualized teaching and learning. Despite the diverse cultural hybridities of these countries, science education is disconnected from the daily lives of the majority of their populations, serving inequitably the academic Western-oriented aspirations of an elite group who are "living hybridity but talking scientism." The discussants explore their autobiographies to reveal core cultural values and beliefs grounded in their non-Western traditions and worldviews but which are in conflict with the Western Modern Worldview (WMW) and thus have no legitimate role in the standard school/college science classroom. They reflect on their hybrid cultural identities and reveal the interplay of multiple selves grounded in both the WMW and non-WMWs and existing in a dialectical tension of managed contradiction in a Third Space. They argue for dialectical logic to illuminate a Third Space wherein students of science education may be empowered to challenge hegemonies of cultural reproduction and examine reflexively their own identities, coming to recognize and reconcile their core cultural beliefs with those of Western modern science, thereby dissipating otherwise strongly delineated cultural borders.
-
Kim, Sun Young; Nehm, Ross H.
2011-01-01
Despite a few international comparisons of the evolutionary beliefs of the general public, comparatively less research has focused on science teachers. Cross-cultural studies offer profitable opportunities for exploring the interactions among knowledge and belief variables in regard to evolution in different socio-cultural contexts. We investigated the evolutionary worldviews of pre-service science teachers from Asia (specifically South Korea), a region often excluded from international comparisons. We compared Korean and American science teachers': (1) understandings of evolution and the nature of science, and (2) acceptance of evolution in order to elucidate how knowledge and belief relationships are manifested in different cultural contexts. We found that Korean science teachers exhibited 'moderate' evolutionary acceptance levels comparable to or lower than American science teacher samples. Gender was significantly related to Korean teachers' evolution content knowledge and acceptance of evolution, with female Christian biology teachers displaying the lowest values on all measures. Korean science teachers' understandings of nature of science were significantly related to their acceptance and understanding of evolution; this relationship appears to transcend cultural boundaries. Our new data on Korean teachers, combined with studies from more than 20 other nations, expose the global nature of science teacher ambivalence or antipathy toward evolutionary knowledge.
-
Principles of cancer cell culture.
Cree, Ian A
2011-01-01
The basics of cell culture are now relatively common, though it was not always so. The pioneers of cell culture would envy our simple access to manufactured plastics, media and equipment for such studies. The prerequisites for cell culture are a well lit and suitably ventilated laboratory with a laminar flow hood (Class II), CO(2) incubator, benchtop centrifuge, microscope, plasticware (flasks and plates) and a supply of media with or without serum supplements. Not only can all of this be ordered easily over the internet, but large numbers of well-characterised cell lines are available from libraries maintained to a very high standard allowing the researcher to commence experiments rapidly and economically. Attention to safety and disposal is important, and maintenance of equipment remains essential. This chapter should enable researchers with little prior knowledge to set up a suitable laboratory to do basic cell culture, but there is still no substitute for experience within an existing well-run laboratory.
-
Perceptions and Practices of Culturally Relevant Science Teaching in American Indian Classrooms
Nam, Younkyeong; Roehrig, Gillian; Kern, Anne; Reynolds, Bree
2013-01-01
This study explores the perceptions of culturally relevant science teaching of 35 teachers of American Indian students. These teachers participated in professional development designed to help them better understand climate change science content and teaching climate change using both Western science and traditional and cultural knowledge. Teacher…
-
sprotocols
2014-01-01
### Materials 1. Glass culture tubes with metal caps and labels - Growth medium, from media room or customized - Glass pipette tubes - Parafilm ### Equipment 1. Vortexer - Fireboy or Bunsen burner - Motorized pipette - Micropipettes and sterile tips ### Procedure For a typical liquid culture, use 5 ml of appropriate medium. The amount in each tube does not have to be exact if you are just trying to culture cells for their precious DNA. 1. Streak an a...
-
Replication of cultured lung epithelial cells
International Nuclear Information System (INIS)
Guzowski, D.; Bienkowski, R.
1986-01-01
The authors have investigated the conditions necessary to support replication of lung type 2 epithelial cells in culture. Cells were isolated from mature fetal rabbit lungs (29d gestation) and cultured on feeder layers of mitotically inactivated 3T3 fibroblasts. The epithelial nature of the cells was demonstrated by indirect immunofluorescent staining for keratin and by polyacid dichrome stain. Ultrastructural examination during the first week showed that the cells contained myofilaments, microvilli and lamellar bodies (markers for type 2 cells). The following changes were observed after the first week: increase in cell size; loss of lamellar bodies and appearance of multivesicular bodies; increase in rough endoplasmic reticulum and golgi; increase in tonafilaments and well-defined junctions. General cell morphology was good for up to 10 wk. Cells cultured on plastic surface degenerated after 1 wk. Cell replication was assayed by autoradiography of cultures exposed to ( 3 H)-thymidine and by direct cell counts. The cells did not replicate during the first week; however, between 2-10 wk the cells incorporated the label and went through approximately 6 population doublings. They have demonstrated that lung alveolar epithelial cells can replicate in culture if they are maintained on an appropriate substrate. The coincidence of ability to replicate and loss of markers for differentiation may reflect the dichotomy between growth and differentiation commonly observed in developing systems
-
Reversible gelling culture media for in-vitro cell culture in three-dimensional matrices
An, Yuehuei H.; Mironov, Vladimir A.; Gutowska, Anna
2000-01-01
A gelling cell culture medium useful for forming a three dimensional matrix for cell culture in vitro is prepared by copolymerizing an acrylamide derivative with a hydrophilic comonomer to form a reversible (preferably thermally reversible) gelling linear random copolymer in the form of a plurality of linear chains having a plurality of molecular weights greater than or equal to a minimum gelling molecular weight cutoff, mixing the copolymer with an aqueous solvent to form a reversible gelling solution and adding a cell culture medium to the gelling solution to form the gelling cell culture medium. Cells such as chondrocytes or hepatocytes are added to the culture medium to form a seeded culture medium, and temperature of the medium is raised to gel the seeded culture medium and form a three dimensional matrix containing the cells. After propagating the cells in the matrix, the cells may be recovered by lowering the temperature to dissolve the matrix and centrifuging.
-
Integration of Culturally Relevant Pedagogy Into the Science Learning Progression Framework
Bernardo, Cyntra
This study integrated elements of culturally relevant pedagogy into a science learning progression framework, with the goal of enhancing teachers' cultural knowledge and thereby creating better teaching practices in an urban public high school science classroom. The study was conducted using teachers, an administrator, a science coach, and students involved in science courses in public high school. Through a qualitative intrinsic case study, data were collected and analyzed using traditional methods. Data from primary participants (educators) were analyzed through identification of big ideas, open coding, and themes. Through this process, patterns and emergent ideas were reported. Outcomes of this study demonstrated that educators lack knowledge about research-based academic frameworks and multicultural education strategies, but benefit through institutionally-based professional development. Students from diverse cultures responded positively to culturally-based instruction. Their progress was further manifested in better communication and discourse with their teacher and peers, and increased academic outcomes. This study has postulated and provided an exemplar for science teachers to expand and improve multicultural knowledge, ultimately transferring these skills to their pedagogical practice.
-
Traditional and Modern Cell Culture in Virus Diagnosis.
Hematian, Ali; Sadeghifard, Nourkhoda; Mohebi, Reza; Taherikalani, Morovat; Nasrolahi, Abbas; Amraei, Mansour; Ghafourian, Sobhan
2016-04-01
Cell cultures are developed from tissue samples and then disaggregated by mechanical, chemical, and enzymatic methods to extract cells suitable for isolation of viruses. With the recent advances in technology, cell culture is considered a gold standard for virus isolation. This paper reviews the evolution of cell culture methods and demonstrates why cell culture is a preferred method for identification of viruses. In addition, the advantages and disadvantages of both traditional and modern cell culture methods for diagnosis of each type of virus are discussed. Detection of viruses by the novel cell culture methods is considered more accurate and sensitive. However, there is a need to include some more accurate methods such as molecular methods in cell culture for precise identification of viruses.
-
Labouta, Hagar Ibrahim; Adams, Jennifer Dawn; Cramb, David Thomas
2018-03-01
In this paper we reflect on the article "I am smart enough to study postsecondary science: a critical discourse analysis of latecomers' identity construction in an online forum", by Phoebe Jackson and Gale Seiler (Cult Stud Sci Educ. https://doi.org/10.1007/s11422-017-9818-0). In their article, the authors did a significant amount of qualitative analysis of a discussion on an online forum by four latecomer students with past negative experiences in science education. The students used this online forum as an out-of-class resource to develop a cultural model based on their ability to ask questions together with solidarity as a new optimistic way to position themselves in science. In this forum, we continue by discussing the identity of marginalized science students in relation to resources available in postsecondary science classes. Recent findings on a successful case of a persistent marginalized science student in spite of prior struggles and failures are introduced. Building on their model and our results, we proposed a new cultural model, emphasizing interaction between inside and outside classroom resources which can further our understanding of the identity of marginalized science students. Exploring this cultural model could better explain drop-outs or engagement of marginalized science students to their study. We, then, used this model to reflect on both current traditional and effective teaching and learning practices truncating or re-enforcing relationships of marginalized students with the learning environment. In this way, we aim to further the discussion initiated by Jackson and Seiler and offer possible frameworks for future research on the interactions between marginalized students with past low achievements and other high and mid achieving students, as well as other interactions between resources inside and outside science postsecondary classrooms.
-
Application of cell co-culture system to study fat and muscle cells.
Pandurangan, Muthuraman; Hwang, Inho
2014-09-01
Animal cell culture is a highly complex process, in which cells are grown under specific conditions. The growth and development of these cells is a highly unnatural process in vitro condition. Cells are removed from animal tissues and artificially cultured in various culture vessels. Vitamins, minerals, and serum growth factors are supplied to maintain cell viability. Obtaining result homogeneity of in vitro and in vivo experiments is rare, because their structure and function are different. Living tissues have highly ordered complex architecture and are three-dimensional (3D) in structure. The interaction between adjacent cell types is quite distinct from the in vitro cell culture, which is usually two-dimensional (2D). Co-culture systems are studied to analyze the interactions between the two different cell types. The muscle and fat co-culture system is useful in addressing several questions related to muscle modeling, muscle degeneration, apoptosis, and muscle regeneration. Co-culture of C2C12 and 3T3-L1 cells could be a useful diagnostic tool to understand the muscle and fat formation in animals. Even though, co-culture systems have certain limitations, they provide a more realistic 3D view and information than the individual cell culture system. It is suggested that co-culture systems are useful in evaluating the intercellular communication and composition of two different cell types.
-
Exploring the Impact of Culture- and Language-Influenced Physics on Science Attitude Enhancement
Morales, Marie Paz E.
2016-02-01
"Culture," a set of principles that trace and familiarize human beings within their existential realities, may provide an invisible lens through which reality could be discerned. Critically explored in this study is how culture- and language-sensitive curriculum materials in physics improve Pangasinan learners' attitude toward science. Their cultural preference or profile defined their cultural dimensions, epistemological beliefs, and views on integration of culture and language in the teaching and learning processes. The culture- and language-influenced curriculum materials in physics were heavily influenced by Pangasinan learners' cultural preference or profile. Results of the experimental participants' pretest and posttest on science attitude measure, when compared, showed significant statistical difference. Assessment of science attitude enhancement favored the experimental group over the control group. Qualitative data gathered from postimplementation interviews, focus group discussions, and journal log entries indicated the same trend in favor of the experimental participants. The study yielded that culture and language integration in the teaching and learning processes of physics concepts allowed students to develop positive attitude to science, their culture, and native language.
-
"Two Cultures" Topics for General Studies Science Courses.
Larson, James H.
1982-01-01
Theses proposed in C. P. Snow's book "The Two Cultures," including uncommunicative scientific and literary groups, gap between rich and poor, overpopulation, and nuclear war remain viable topics. Discusses the scientific and literary cultural gap and what can be done in general studies science courses to ameliorate the condition.…
-
Medrano, Jose V; Rombaut, Charlotte; Simon, Carlos; Pellicer, Antonio; Goossens, Ellen
2016-11-01
To study the ability of human spermatogonial stem cells (hSSCs) to proliferate in vitro under mouse spermatogonial stem cell (mSSC) culture conditions. Experimental basic science study. Reproductive biology laboratory. Cryopreserved testicular tissue with normal spermatogenesis obtained from three donors subjected to orchiectomy due to a prostate cancer treatment. Testicular cells used to create in vitro cell cultures corresponding to the following groups: [1] unsorted human testicular cells, [2] differentially plated human testicular cells, and [3] cells enriched with major histocompatibility complex class 1 (HLA - )/epithelial cell surface antigen (EPCAM + ) in coculture with inactivated testicular feeders from the same patient. Analyses and characterization including immunocytochemistry and quantitative reverse-transcription polymerase chain reaction for somatic and germ cell markers, testosterone and inhibin B quantification, and TUNEL assay. Putative hSSCs appeared in singlets, doublets, or small groups of up to four cells in vitro only when testicular cells were cultured in StemPro-34 medium supplemented with glial cell line-derived neurotrophic factor (GDNF), leukemia inhibitory factor (LIF), basic fibroblast growth factor (bFGF), and epidermal growth factor (EGF). Fluorescence-activated cell sorting with HLA - /EPCAM + resulted in an enrichment of 27% VASA + /UTF1 + hSSCs, compared to 13% in unsorted controls. Coculture of sorted cells with inactivated testicular feeders gave rise to an average density of 112 hSSCs/cm 2 after 2 weeks in vitro compared with unsorted cells (61 hSSCs/cm 2 ) and differentially plated cells (49 hSSCS/cm 2 ). However, putative hSSCs rarely stained positive for the proliferation marker Ki67, and their presence was reduced to the point of almost disappearing after 4 weeks in vitro. We found that hSSCs show limited proliferation in vitro under mSSC culture conditions. Coculture of HLA - /EPCAM + sorted cells with testicular
-
Torgan, C. E.; Burge, S. S.; Collinsworth, A. M.; Truskey, G. A.; Kraus, W. E.
2000-01-01
The growth and repair of adult skeletal muscle are due in part to activation of muscle precursor cells, commonly known as satellite cells or myoblasts. These cells are responsive to a variety of environmental cues, including mechanical stimuli. The overall goal of the research is to examine the role of mechanical signalling mechanisms in muscle growth and plasticity through utilisation of cell culture systems where other potential signalling pathways (i.e. chemical and electrical stimuli) are controlled. To explore the effects of decreased mechanical loading on muscle differentiation, mammalian myoblasts are cultured in a bioreactor (rotating cell culture system), a model that has been utilised to simulate microgravity. C2C12 murine myoblasts are cultured on microcarrier beads in a bioreactor and followed throughout differentiation as they form a network of multinucleated myotubes. In comparison with three-dimensional control cultures that consist of myoblasts cultured on microcarrier beads in teflon bags, myoblasts cultured in the bioreactor exhibit an attenuation in differentiation. This is demonstrated by reduced immunohistochemical staining for myogenin and alpha-actinin. Western analysis shows a decrease, in bioreactor cultures compared with control cultures, in levels of the contractile proteins myosin (47% decrease, p < 0.01) and tropomyosin (63% decrease, p < 0.01). Hydrodynamic measurements indicate that the decrease in differentiation may be due, at least in part, to fluid stresses acting on the myotubes. In addition, constraints on aggregate size imposed by the action of fluid forces in the bioreactor affect differentiation. These results may have implications for muscle growth and repair during spaceflight.
-
Politics as Culture: Contribution of Political Science to Democratic Maturity
Directory of Open Access Journals (Sweden)
Ivan Padjen
2014-01-01
Full Text Available The article discusses the contribution of Croatian political science to the development of democracy in Croatia. The focus of the analysis is the concept of culture which author talks about in five steps. In the first step it is understood in the modern key, in the second step as different for nature and in the third as different from society. In the fourth step author differentiates political culture from political economy and political institutions, but in the fifth part there is an attempt to show culture as a fundamental part of politics, policy and polity. On the basis of these insights author shows that the matrix of Croatian political science is more and more devoted to scientific investigation of politics as culture as both study of political culture and as a source of development as politics as culture.
-
Bulletin of Materials Science | Indian Academy of Sciences
Indian Academy of Sciences (India)
Home; Journals; Bulletin of Materials Science; Volume 29; Issue 5 ... Polyester urethane; scaffold; tensile strength; swelling; degradation; cell culture. ... Materials Science Centre, Indian Institute of Technology, Kharagpur 721 302, India; School of Medical Science and Technology, Indian Institute of Technology, Kharagpur ...
-
Renegotiating forensic cultures: between law, science and criminal justice.
Roberts, Paul
2013-03-01
This article challenges stereotypical conceptions of Law and Science as cultural opposites, arguing that English criminal trial practice is fundamentally congruent with modern science's basic epistemological assumptions, values and methods of inquiry. Although practical tensions undeniably exist, they are explicable-and may be neutralised-by paying closer attention to criminal adjudication's normative ideals and their institutional expression in familiar aspects of common law trial procedure, including evidentiary rules of admissibility, trial by jury, adversarial fact-finding, cross-examination and the ethical duties of expert witnesses. Effective partnerships between lawyers and forensic scientists are indispensable for integrating scientific evidence into criminal proceedings, and must be renegotiated between individual practitioners on an on-going basis. Fruitful interdisciplinary collaboration between scholars with a shared interest in forensic science should dispense with reductive cultural stereotypes of Science and Law. Copyright © 2013. Published by Elsevier Ltd.
-
Flow field measurements in the cell culture unit
Walker, Stephen; Wilder, Mike; Dimanlig, Arsenio; Jagger, Justin; Searby, Nancy
2002-01-01
The cell culture unit (CCU) is being designed to support cell growth for long-duration life science experiments on the International Space Station (ISS). The CCU is a perfused loop system that provides a fluid environment for controlled cell growth experiments within cell specimen chambers (CSCs), and is intended to accommodate diverse cell specimen types. Many of the functional requirements depend on the fluid flow field within the CSC (e.g., feeding and gas management). A design goal of the CCU is to match, within experimental limits, all environmental conditions, other than the effects of gravity on the cells, whether the hardware is in microgravity ( micro g), normal Earth gravity, or up to 2g on the ISS centrifuge. In order to achieve this goal, two steps are being taken. The first step is to characterize the environmental conditions of current 1g cell biology experiments being performed in laboratories using ground-based hardware. The second step is to ensure that the design of the CCU allows the fluid flow conditions found in 1g to be replicated from microgravity up to 2g. The techniques that are being used to take these steps include flow visualization, particle image velocimetry (PIV), and computational fluid dynamics (CFD). Flow visualization using the injection of dye has been used to gain a global perspective of the characteristics of the CSC flow field. To characterize laboratory cell culture conditions, PIV is being used to determine the flow field parameters of cell suspension cultures grown in Erlenmeyer flasks on orbital shakers. These measured parameters will be compared to PIV measurements in the CSCs to ensure that the flow field that cells encounter in CSCs is within the bounds determined for typical laboratory experiments. Using CFD, a detailed simulation is being developed to predict the flow field within the CSC for a wide variety of flow conditions, including microgravity environments. Results from all these measurements and analyses of the
-
International Nuclear Information System (INIS)
Fan, Ping; He, Lan; Pu, Dan; Lv, Xiaohong; Zhou, Wenxu; Sun, Yining; Hu, Nan
2011-01-01
Research highlights: → The proliferation of dramatic increased by co-cultured with Sertoli cells. → VEGF receptor-2 expression of ECs was up-regulated by co-cultured with Sertoli cells. → The MHC expression of ECs induced by INF-γ and IL-6, IL-8 and sICAM induced by TNF-α decreased respectively after co-cultured with Sertoli cells. → ECs co-cultured with Sertoli cells also didn't increase the stimulation index of spleen lymphocytes. -- Abstract: The major problem of the application of endothelial cells (ECs) in transplantation is the lack of proliferation and their immunogenicity. In this study, we co-cultured ECs with Sertoli cells to monitor whether Sertoli cells can influence the proliferation and immunogenicity of co-cultured ECs. Sertoli cells were isolated from adult testicular tissue. ECs were divided into the control group and the experimental group, which included three sub-groups co-cultured with 1 x 10 3 , 1 x 10 4 or 1 x 10 5 cell/ml of Sertoli cells. The growth and proliferation of ECs were observed microscopically, and the expression of vascular endothelial growth factor (VEGF) receptor-2 (KDR) was examined by Western blotting. In another experiment, ECs were divided into the control group, the single culture group and the co-culture group with the optimal concentration of Sertoli cells. After INF-γ and TNF-α were added to the culture medium, MHC II antigen expression was detected by immunofluorescence staining and western blotting; interleukin (IL)-6, IL-8 and soluble intercellular adhesion molecule (sICAM) were measured in the culture medium by ELISA. We demonstrated that 1 x 10 4 cell/ml Sertoli cells promoted the proliferation of co-cultured ECs more dramatically than that in other groups (P 4 cell/ml of the Sertoli cells was most effective in the up-regulation of KDR expression in the co-cultured ECs (P < 0.05). Sertoli cells can effectively suppress INF-γ-induced MHC II antigen expression in co-cultured ECs compared with single
-
SCIENCE WHERE CULTURE MATTERS: A NEO-CLASSICAL ...
Indian Academy of Sciences (India)
SCIENCE WHERE CULTURE MATTERS: A NEO-CLASSICAL APPROACH TO EXPLORE UNTAPPED BACTERIAL DIVERSITY. MILIND WATVE; Dept of Microbiology, Abasaheb Garware College, Pune. www.culturematters.org; * Life Research Foundation, Pune; * Evolvus Biotech Pvt. Ltd.,Pune ...
-
Andreeva, Elena; Andrianova, Irina; Rylova, Julia; Gornostaeva, Aleksandra; Bobyleva, Polina; Buravkova, Ludmila
2015-08-01
The impact of culture conditions and interaction with activated peripheral blood mononuclear cells on the interleukin (IL) gene expression profile and proinflammatory IL-6 and IL-8 production by adipose-derived stromal cells (ASCs) was investigated. A microarray analysis revealed a wide range of IL genes either under standard (20%) or hypoxic (5%) O2 concentrations, some highly up-regulated at hypoxia. IL-6 and IL-8 production was inversely dependent on cell culture density. In early (first-third) passages, IL-6 and IL-8 concentration was higher at 20% O2 and in late (8th-12th) passages under 5% O2. Interaction between ASCs and mononuclear cells in indirect setting was accompanied with a significant decrease of IL-6 and did not result in the elevation of IL-8 concentration. Thereby, the production of proinflammatory interleukins (IL-6 and IL-8) may be affected by the ASC intrinsic features (density in culture, and duration of expansion), as well as by microenvironmental factors, such as hypoxia and the presence of blood-borne cells. These data are important for elucidating ASC paracrine activity regulation in vitro. They would also be on demand for optimisation of the cell therapy protocols, based on the application of ASC biologically active substances. SIGNIFICANCE PARAGRAPH: Ex vivo expansion is widely used for increasing the number of adipose-derived stromal cells (ASCs) and improving of their quality. The present study was designed to elucidate the particular factors influencing the interleukin production in ASCs. The presented data specified the parameters (i.e. cell density, duration of cultivation, hypoxia, etc.) that should be taken in mind when ASCs are intended to be used in protocols implying their paracrine activity. These data would be of considerable interest for researchers and clinicians working in the biomedical science. Copyright © 2015 John Wiley & Sons, Ltd.
-
Garba, Abubakar; Acar, Delphine D; Roukaerts, Inge D M; Desmarets, Lowiese M B; Devriendt, Bert; Nauwynck, Hans J
2017-09-01
Mesenchymal cells are multipotent stromal cells with self-renewal, differentiation and immunomodulatory capabilities. We aimed to develop a co-culture model for differentiating hematopoietic cells on top of immortalized mesenchymal cells for studying interactions between hematopoietic and mesenchymal cells, useful for adequately exploring the therapeutic potential of mesenchymal cells. In this study, we investigated the survival, proliferation and differentiation of porcine red bone marrow hematopoietic cells co-cultured with immortalized porcine bone marrow mesenchymal cells for a period of five weeks. Directly after collection, primary porcine bone marrow mesenchymal cells adhered firmly to the bottom of the culture plates and showed a fibroblast-like appearance, one week after isolation. Upon immortalization, porcine bone marrow mesenchymal cells were continuously proliferating. They were positive for simian virus 40 (SV40) large T antigen and the mesenchymal cell markers CD44 and CD55. Isolated red bone marrow cells were added to these immortalized mesenchymal cells. Five weeks post-seeding, 92±6% of the red bone marrow hematopoietic cells were still alive and their number increased 3-fold during five weekly subpassages on top of the immortalized mesenchymal cells. The red bone marrow hematopoietic cells were originally small and round; later, the cells increased in size. Some of them became elongated, while others remained round. Tiny dendrites appeared attaching hematopoietic cells to the underlying immortalized mesenchymal cells. Furthermore, weekly differential-quick staining of the cells indicated the presence of monoblasts, monocytes, macrophages and lymphocytes in the co-cultures. At three weeks of co-culture, flow cytometry analysis showed an increased surface expression of CD172a, CD14, CD163, CD169, CD4 and CD8 up to 37±0.8%, 40±8%, 41±4%, 23±3% and 19±5% of the hematopoietic cells, respectively. In conclusion, continuous mesenchymal cell
-
3D Cell Culture in Alginate Hydrogels
Directory of Open Access Journals (Sweden)
Therese Andersen
2015-03-01
Full Text Available This review compiles information regarding the use of alginate, and in particular alginate hydrogels, in culturing cells in 3D. Knowledge of alginate chemical structure and functionality are shown to be important parameters in design of alginate-based matrices for cell culture. Gel elasticity as well as hydrogel stability can be impacted by the type of alginate used, its concentration, the choice of gelation technique (ionic or covalent, and divalent cation chosen as the gel inducing ion. The use of peptide-coupled alginate can control cell–matrix interactions. Gelation of alginate with concomitant immobilization of cells can take various forms. Droplets or beads have been utilized since the 1980s for immobilizing cells. Newer matrices such as macroporous scaffolds are now entering the 3D cell culture product market. Finally, delayed gelling, injectable, alginate systems show utility in the translation of in vitro cell culture to in vivo tissue engineering applications. Alginate has a history and a future in 3D cell culture. Historically, cells were encapsulated in alginate droplets cross-linked with calcium for the development of artificial organs. Now, several commercial products based on alginate are being used as 3D cell culture systems that also demonstrate the possibility of replacing or regenerating tissue.
-
Kant and the development of the human and cultural sciences.
Makkreel, Rudolf A
2008-12-01
Starting with Kant's doubts about psychology as a natural science capable of explaining human behavior, several alternative attempts to conceive of human life, culture and history are examined. Kant proposes an anthropology that will be a commonly useful human science rather than a universally valid natural science. This anthropology relates to philosophy as a mode of world-cognition. Special attention is given to how Kant's theory of right can help define our appropriate place in a communal world. The different ways in which Wilhelm Dilthey and Hermann Cohen respond to Kant's idea of legitimate appropriation are also considered. The various tasks that descriptive elucidation, explanation, reflective understanding, characterization and interpretation can perform for the human and cultural sciences are examined throughout the essay.
-
Astrobiology in culture: the search for extraterrestrial life as "science".
Billings, Linda
2012-10-01
This analysis examines the social construction of authority, credibility, and legitimacy for exobiology/astrobiology and, in comparison, the search for extraterrestrial intelligence (SETI), considering English-language conceptions of these endeavors in scientific culture and popular culture primarily in the United States. The questions that define astrobiology as a scientific endeavor are multidisciplinary in nature, and this endeavor is broadly appealing to public audiences as well as to the scientific community. Thus, it is useful to examine astrobiology in culture-in scientific culture, official culture, and popular culture. A researcher may explore science in culture, science as culture, by analyzing its rhetoric, the primary means that people use to construct their social realities-their cultural environment, as it were. This analysis follows this path, considering scientific and public interest in astrobiology and SETI and focusing on scientific and official constructions of the two endeavors. This analysis will also consider whether and how scientific and public conceptions of astrobiology and SETI, which are related but at the same time separate endeavors, converge or diverge and whether and how these convergences or divergences affect the scientific authority, credibility, and legitimacy of these endeavors.
-
Advances in cell culture: anchorage dependence
Merten, Otto-Wilhelm
2015-01-01
Anchorage-dependent cells are of great interest for various biotechnological applications. (i) They represent a formidable production means of viruses for vaccination purposes at very large scales (in 1000–6000 l reactors) using microcarriers, and in the last decade many more novel viral vaccines have been developed using this production technology. (ii) With the advent of stem cells and their use/potential use in clinics for cell therapy and regenerative medicine purposes, the development of novel culture devices and technologies for adherent cells has accelerated greatly with a view to the large-scale expansion of these cells. Presently, the really scalable systems—microcarrier/microcarrier-clump cultures using stirred-tank reactors—for the expansion of stem cells are still in their infancy. Only laboratory scale reactors of maximally 2.5 l working volume have been evaluated because thorough knowledge and basic understanding of critical issues with respect to cell expansion while retaining pluripotency and differentiation potential, and the impact of the culture environment on stem cell fate, etc., are still lacking and require further studies. This article gives an overview on critical issues common to all cell culture systems for adherent cells as well as specifics for different types of stem cells in view of small- and large-scale cell expansion and production processes. PMID:25533097
-
Goldston, M. Jenice; Nichols, Sharon
2009-04-01
This study situated in a Southern resegregated Black middle school involved four Black teachers and two White science educators’ use of photonarratives to envision culturally relevant science pedagogy. Two questions guided the study: (1) What community referents are important for conceptualizing culturally relevant practices in Black science classrooms? and (2) How do teachers’ photonarratives serve to open conversations and notions of culturally relevant science practices? The research methodologically drew upon memory-work, Black feminism, critical theory, visual methodology, and narrative inquiry as “portraiture.” Issues of positionality and identity proved to be central to this work, as three luminaries portray Black teachers’ insights about supports and barriers to teaching and learning science. The community referents identified were associated with church and its oral traditions, inequities of the market place in meeting their basic human needs, and community spaces.
-
Exploring the development of a cultural care framework for European caring science.
Albarran, John; Rosser, Elizabeth; Bach, Shirley; Uhrenfeldt, Lisbeth; Lundberg, Pranee; Law, Kate
2011-01-01
The aim of this paper is to discuss the development of a cultural care framework that seeks to inform and embrace the philosophical ideals of caring science. Following a review of the literature that identified a lack of evidence of an explicit relationship between caring science and cultural care, a number of well-established transcultural care frameworks were reviewed. Our purpose was to select one that would resonate with underpinning philosophical values of caring science and that drew on criteria generated by the European Academy of Caring Science members. A modified framework based on the work of Giger and Davidhizar was developed as it embraced many of the values such as humanism that are core to caring science practice. The proposed caring science framework integrates determinants of cultural lifeworld-led care and seeks to provide clear directions for humanizing the care of individuals. The framework is offered to open up debate and act as a platform for further academic enquiry.
-
Baron, Alex; Chen, Hsiao-Lan Sharon
2012-03-01
Worldwide proliferation of pedagogical innovations creates expanding potential in the field of science education. While some teachers effectively improve students' scientific learning, others struggle to achieve desirable student outcomes. This study explores a Taiwanese science teacher's ability to effectively enhance her students' science learning. The authors visited a Taipei city primary school class taught by an experienced science teacher during a 4-week unit on astronomy, with a total of eight, 90-minute periods. Research methods employed in this study included video capture of each class as well as reflective interviews with the instructor, eliciting the teacher's reflection upon both her pedagogical choices and the perceived results of these choices. We report that the teacher successfully teaches science by creatively diverging from culturally generated educational expectations. Although the pedagogical techniques and ideas enumerated in the study are relevant specifically to Taiwan, creative cultural divergence might be replicated to improve science teaching worldwide.
-
Influence of Culture and Language Sensitive Physics on Science Attitude Enhancement
Morales, Marie Paz E.
2015-01-01
The study critically explored how culture and language sensitive curriculum materials in physics improve Pangasinan learners' attitude towards science. Their cultural dimensions, epistemological beliefs, and views on integration of culture and language in the teaching and learning process determined their cultural preference or profile. Design and…
-
Suspension culture of pluripotent stem cells: effect of shear on stem cell fate.
Keller, Kevin C; Rodrigues, Beatriz; zur Nieden, Nicole I
2014-01-01
Despite significant promise, the routine usage of suspension cell culture to manufacture stem cell-derived differentiated cells has progressed slowly. Suspension culture is an innovative way of either expanding or differentiating cells and sometimes both are combined into a single bioprocess. Its advantages over static 2D culturing include a homogeneous and controllable culture environment and producing a large quantity of cells in a fraction of time. This feature makes suspension cell culture ideal for use in stem cell research and eventually ideal in the large-scale production of differentiated cells for regenerative medicine. Because of their tremendous differentiation capacities and unlimited growth properties, pluripotent stem cells (PSCs) in particular are considered potential sources for future cell-replacement therapies. Currently, expansion of PSCs is accomplished in 2D, which only permits a limited amount of cell growth per culture flask before cells need to be passaged. However, before stem cells can be applied clinically, several aspects of their expansion, such as directed growth, but also differentiation, need to be better controlled. This review will summarize recent advantages in suspension culture of PSCs, while at the same time highlighting current challenges.
-
Morrow, Cherilynn A.
1993-01-01
The visceral appeal of space science and exploration is a very powerful emotional connection to a very large and diverse collection of people, most of whom have little or no perspective about what it means to do science and engineering. Therein lies the potential of space for a substantially enhanced positive impact on culture through education. This essay suggests that through engaging more of the space research and development community in enabling unique and 'meaningful educational experiences' for educators and students at the pre-collegiate levels, space science and exploration can amplify its positive feedback on society and act as an important medium for cultural transformation to greater science literacy. I discuss the impact of space achievements on people and define what is meant by a 'meaningful educational experience,' all of which points to the need for educators and students to be closer to the practice of real science. I offer descriptions of two nascent science education programs associated with NASA which have the needed characteristics for providing meaningful experiences that can cultivate greater science literacy. Expansion of these efforts and others like it will be needed to have the desired impact on culture, but I suggest that the potential for the needed resources is there in the scientific research communities. A society in which more people appreciate and understand science and science methods would be especially conducive to human progress in space and on Earth.
-
Controlling the diversity of cell populations in a stem cell culture
Heo, Inha; Clevers, Hans
2015-01-01
Culturing intestinal stem cells into 3D organoids results in heterogeneous cell populations, reflecting the in vivo cell type diversity. In a recent paper published in Nature, Wang et al. established a culture condition for a highly homogeneous population of intestinal stem cells.
-
Mammalian Cell Culture Simplified.
Moss, Robert; Solomon, Sondra
1991-01-01
A tissue culture experiment that does not require elaborate equipment and that can be used to teach sterile technique, the principles of animal cell line maintenance, and the concept of cell growth curves is described. The differences between cancerous and normal cells can be highlighted. The procedure is included. (KR)
-
Reiners, Christiane S.; Bliersbach, Markus; Marniok, Karl
2017-07-01
Understanding Nature of Science (NOS) is a central component of scientific literacy, which is agreed upon internationally, and consequently has been a major educational goal for many years all over the globe. In order to justify the promotion of an adequate understanding of NOS, educators have developed several arguments, among them the cultural argument. But what is behind this argument? In order to answer this question, C. P. Snow's vision of two cultures was used as a starting point. In his famous Rede Lecture from 1959, he complained about a wide gap between the arts and humanities on the one hand and sciences on the other hand. While the representatives of the humanities refer to themselves as real intellectuals, the scientists felt rather ignored as a culture, despite the fact that their achievements had been so important for Western society. Thus, Snow argued that as these intellectual cultures were completely different from each other, a mutual understanding was impossible. The first European Regional IHPST Conference took up the cultural view on science again. Thus, the topic of the conference "Science as Culture in the European Context" encouraged us to look at the two cultures and to figure out possibilities to bridge the gap between them in chemistry teacher education. For this reason, we put together three studies—one theoretical and two independent research projects (one dealing with creativity in science, the other with scientific laws and theories) which contribute to our main research field (promoting an understanding of NOS)—in order to address the cultural argument for understanding science from an educational point of view. Among the consented tenets of what understanding NOS implies in an educational context, there are aspects which are associated mainly with the humanities, like the tentativeness of knowledge, creativity, and social tradition, whereas others seem to have a domain-specific meaning, like empirical evidence, theories and laws
-
Rabbit uterine epithelial cells: Co-culture with spermatozoa
International Nuclear Information System (INIS)
Boice, M.L.
1988-01-01
A primary culture of rabbit uterine epithelial cells was established and their effects on sperm function were examined in vitro. Epithelial cells were isolated from uteri of estrous rabbits and cultured on floating collagen gels in phenol red-free medium supplemented with 5% fetal bovine serum. Light microscopy and keratin staining showed that the epithelial cell population established in culture had morphological characteristics similar to that seen in the intact endometrium. Cells were cultured with 3 H-leucine and uptake of label by cells and its incorporation into cellular and secretory proteins determined. When compared to cells cultured for 24-48 h, incorporation of label into cellular protein was lower at 72-96 h, but secretion increased. Estradiol 17-β did not affect label uptake or incorporation, but did enhance proliferation of cells as judged by total DNA content of the cell population. Analysis of proteins in media by sodium dodecyl sulfate polyacrylamide gel electrophoresis and fluorography suggested that epithelial and stromal cells synthesis proteins that may be secretory in nature during 72-96 h culture. Twenty-nine to thirty-one h after initiation of epithelial cultures, 1-2 x 10 6 sperm were co-incubated with cells and sperm viability, motility, loss of acrosome and fertilizing ability determined
-
X-ray microanalysis of single and cultured cells
International Nuclear Information System (INIS)
Wroblewski, J.; Roomans, G.M.
1984-01-01
X-ray microanalysis of single or cultured cells is often a useful alternative or complement to the analysis of the corresponding tissue. It also allows the analysis of individual cells in a cell population. Preparation for X-ray microanalysis poses a number of typical problems. Suspensions of single cells can be prepared by either of two pathways: (1) washing - mounting - drying, or (2) centrifugation - freezing or fixation - sectioning. The washing step in the preparation of single or cultured cells presents the most severe problems. Cultured cells are generally grown on a substrate that is compatible with both the analysis and the culture, washed and dried. In some cases, sectioning of cultured cell monolayers has been performed. Special problems in quantitative analysis occur in those cases where the cells are analyzed on a thick substrate, since the substrate contributes to the spectral background
-
[Psychiatry as cultural science: considerations following Max Weber].
Bormuth, M
2010-11-01
Psychiatry can be seen as a natural and cultural science. According to this the postulate of freedom is its strong value judgment. Since the times of enlightenment it has been described metaphorically by the myth of the expulsion from Paradise. Following Max Weber and Wilhelm Dilthey, Karl Jaspers has introduced this perspective into psychiatry. His strict dichotomy between explaining and understanding has later been critically revised by Werner Janzarik and Hans Heimann. Their concepts of structure dynamic, of pathography and of anthropology are closer to Max Weber who connected natural and cultural sciences in a much stronger way. Especially the pathographic example of Nietzsche allows to demonstrate the differences between Jaspers and the later psychopathologists of the Heidelberg and Tübingen schools.
-
Mapping epistemic cultures and learning potential of participants in citizen science projects.
Vallabh, Priya; Lotz-Sisitka, Heila; O'Donoghue, Rob; Schudel, Ingrid
2016-06-01
The ever-widening scope and range of global change and interconnected systemic risks arising from people-environment relationships (social-ecological risks) appears to be increasing concern among, and involvement of, citizens in an increasingly diversified number of citizen science projects responding to these risks. We examined the relationship between epistemic cultures in citizen science projects and learning potential related to matters of concern. We then developed a typology of purposes and a citizen science epistemic-cultures heuristic and mapped 56 projects in southern Africa using this framework. The purpose typology represents the range of knowledge-production purposes, ranging from laboratory science to social learning, whereas the epistemic-cultures typology is a relational representation of scientist and citizen participation and their approach to knowledge production. Results showed an iterative relationship between matters of fact and matters of concern across the projects; the nexus of citizens' engagement in knowledge-production activities varied. The knowledge-production purposes informed and shaped the epistemic cultures of all the sampled citizen science projects, which in turn influenced the potential for learning within each project. Through a historical review of 3 phases in a long-term river health-monitoring project, we found that it is possible to evolve the learning curve of citizen science projects. This evolution involved the development of scientific water monitoring tools, the parallel development of pedagogic practices supporting monitoring activities, and situated engagement around matters of concern within social activism leading to learning-led change. We conclude that such evolutionary processes serve to increase potential for learning and are necessary if citizen science is to contribute to wider restructuring of the epistemic culture of science under conditions of expanding social-ecological risk. © 2016 Society for
-
Particle Trajectories in Rotating Wall Cell Culture Devices
Ramachandran N.; Downey, J. P.
1999-01-01
Cell cultures are extremely important to the medical community since such cultures provide an opportunity to perform research on human tissue without the concerns inherent in experiments on individual humans. Development of cells in cultures has been found to be greatly influenced by the conditions of the culture. Much work has focused on the effect of the motions of cells in the culture relative to the solution. Recently rotating wall vessels have been used with success in achieving improved cellular cultures. Speculation and limited research have focused on the low shear environment and the ability of rotating vessels to keep cells suspended in solution rather than floating or sedimenting as the primary reasons for the improved cellular cultures using these devices. It is widely believed that the cultures obtained using a rotating wall vessel simulates to some degree the effect of microgravity on cultures. It has also been speculated that the microgravity environment may provide the ideal acceleration environment for culturing of cellular tissues due to the nearly negligible levels of sedimentation and shear possible. This work predicts particle trajectories of cells in rotating wall vessels of cylindrical and annular design consistent with the estimated properties of typical cellular cultures. Estimates of the shear encountered by cells in solution and the interactions with walls are studied. Comparisons of potential experiments in ground and microgravity environments are performed.
-
Cultural, Social and Political Perspectives in Science Education
DEFF Research Database (Denmark)
conditions and contexts in science education. The different chapters review debates and research in teacher education, school teaching and learning including when external stakeholders are involved. Even though the chapters are contextualized in Nordic settings there will be similarities and parallels...... that will be informative to the international science education research community.......This book presents a collection of critical thinking that concern cultural, social and political issues for science education in the Nordic countries. The chapter authors describe specific scenarios to challenge persisting views, interrogate frameworks and trouble contemporary approaches...
-
Usability and Applicability of Microfluidic Cell Culture Systems
DEFF Research Database (Denmark)
Hemmingsen, Mette
possibilities for, for example, precise control of the chemical environment, 3D cultures, controlled co-culture of different cell types or automated, individual control of up to 96 cell culture chambers in one integrated system. Despite the great new opportunities to perform novel experimental designs......Microfluidic cell culture has been a research area with great attention the last decade due to its potential to mimic the in vivo cellular environment more closely compared to what is possible by conventional cell culture methods. Many exciting and complex devices have been presented providing......, these devices still lack general implementation into biological research laboratories. In this project, the usability and applicability of microfluidic cell culture systems have been investigated. The tested systems display good properties regarding optics and compatibility with standard laboratory equipment...
-
Science-Based Thematic Cultural Art Learning in Primary School (2013 Curriculum
Directory of Open Access Journals (Sweden)
Warih Handayaningrum
2016-12-01
Full Text Available This study is aimed at discussing the development result of thematic cultural art subject’s learning material based on science for primary school (2013 curriculum. This study is expected to inspire teacher to develop learning material that may explore artworks exist in our living environment (based on the context of children’s environment. This study applies steps in developmental research collaboration by Borg & Gall (1989 and Puslitjaknov (2008 to create the product. The development stages comprise observation in several primary schools in Surabaya, Gresik, and Sidoarjo that has implemented 2013 curriculum that is followed up by stages of development. Furthermore, prototype of cultural and art thematic learning material development results are verified by learning material experts, material expert, primary school teacher, and revised afterwards. The result of this research development is a set of teacher and student books. Science-based cultural art here means cultural art learning as the main medium to introduce local culture products (music, drawing, dance, and drama by integrating mathematics, sciences, Bahasa Indonesia, and local language subjects. Cultural art products in the form of dance, music, drawing, dramas will help children to understand a simple mathematical concept, such as: two-dimensional figure, geometry, comparing or estimating longer-shorter, smaller-bigger, or more-less.
-
Good Cell Culture Practice for stem cells and stem-cell-derived models.
Pamies, David; Bal-Price, Anna; Simeonov, Anton; Tagle, Danilo; Allen, Dave; Gerhold, David; Yin, Dezhong; Pistollato, Francesca; Inutsuka, Takashi; Sullivan, Kristie; Stacey, Glyn; Salem, Harry; Leist, Marcel; Daneshian, Mardas; Vemuri, Mohan C; McFarland, Richard; Coecke, Sandra; Fitzpatrick, Suzanne C; Lakshmipathy, Uma; Mack, Amanda; Wang, Wen Bo; Yamazaki, Daiju; Sekino, Yuko; Kanda, Yasunari; Smirnova, Lena; Hartung, Thomas
2017-01-01
The first guidance on Good Cell Culture Practice (GCCP) dates back to 2005. This document expands this to include aspects of quality assurance for in vitro cell culture focusing on the increasingly diverse cell types and culture formats used in research, product development, testing and manufacture of biotechnology products and cell-based medicines. It provides a set of basic principles of best practice that can be used in training new personnel, reviewing and improving local procedures, and helping to assure standard practices and conditions for the comparison of data between laboratories and experimentation performed at different times. This includes recommendations for the documentation and reporting of culture conditions. It is intended as guidance to facilitate the generation of reliable data from cell culture systems, and is not intended to conflict with local or higher level legislation or regulatory requirements. It may not be possible to meet all recommendations in this guidance for practical, legal or other reasons. However, when it is necessary to divert from the principles of GCCP, the risk of decreasing the quality of work and the safety of laboratory staff should be addressed and any conclusions or alternative approaches justified. This workshop report is considered a first step toward a revised GCCP 2.0.
-
Long-term maintenance of human induced pluripotent stem cells by automated cell culture system.
Konagaya, Shuhei; Ando, Takeshi; Yamauchi, Toshiaki; Suemori, Hirofumi; Iwata, Hiroo
2015-11-17
Pluripotent stem cells, such as embryonic stem cells and induced pluripotent stem (iPS) cells, are regarded as new sources for cell replacement therapy. These cells can unlimitedly expand under undifferentiated conditions and be differentiated into multiple cell types. Automated culture systems enable the large-scale production of cells. In addition to reducing the time and effort of researchers, an automated culture system improves the reproducibility of cell cultures. In the present study, we newly designed a fully automated cell culture system for human iPS maintenance. Using an automated culture system, hiPS cells maintained their undifferentiated state for 60 days. Automatically prepared hiPS cells had a potency of differentiation into three germ layer cells including dopaminergic neurons and pancreatic cells.
-
High-Throughput Cancer Cell Sphere Formation for 3D Cell Culture.
Chen, Yu-Chih; Yoon, Euisik
2017-01-01
Three-dimensional (3D) cell culture is critical in studying cancer pathology and drug response. Though 3D cancer sphere culture can be performed in low-adherent dishes or well plates, the unregulated cell aggregation may skew the results. On contrary, microfluidic 3D culture can allow precise control of cell microenvironments, and provide higher throughput by orders of magnitude. In this chapter, we will look into engineering innovations in a microfluidic platform for high-throughput cancer cell sphere formation and review the implementation methods in detail.
-
Andria, B.; Bracco, A.; Cirino, G.; Chamuleau, R. A. F. M.
2010-01-01
In the last 15 years many different liver cell culture devices, consisting of functional liver cells and artificial materials, have been developed. They have been devised for numerous different applications, such as temporary organ replacement (a bridge to liver transplantation or native liver
-
Developing safety culture-rocket science or common sense?
International Nuclear Information System (INIS)
Mahn, J.A.
1998-01-01
Despite evidence of significant management contributions to the causes of major accidents, recent events at Millstone Nuclear Power Station in the US and Ontario Hydro in Canada might lead one to conclude that the significance of safety culture, and the role of management in developing and maintaining an appropriate safety culture, is either not being understood or not being taken serious as integral to the safe operation of some complex, high-reliability operations. It is the purpose of this paper to address four aspects of management that are particularly important to safety culture, and to illustrate how development of an appropriate safety culture is more a matter of common sense than rocket science
-
Developing safety culture-rocket science or common sense?
Energy Technology Data Exchange (ETDEWEB)
Mahn, J.A.
1998-08-01
Despite evidence of significant management contributions to the causes of major accidents, recent events at Millstone Nuclear Power Station in the US and Ontario Hydro in Canada might lead one to conclude that the significance of safety culture, and the role of management in developing and maintaining an appropriate safety culture, is either not being understood or not being taken serious as integral to the safe operation of some complex, high-reliability operations. It is the purpose of this paper to address four aspects of management that are particularly important to safety culture, and to illustrate how development of an appropriate safety culture is more a matter of common sense than rocket science.
-
Characterization of glucocerebrosidase in peripheral blood cells and cultured blastoid cells
Aerts, J. M.; Heikoop, J.; van Weely, S.; Donker-Koopman, W. E.; Barranger, J. A.; Tager, J. M.; Schram, A. W.
1988-01-01
We have characterized glucocerebrosidase in various cell types of peripheral blood of control subjects and in cultured human blastoid cells. The intracellular level of glucocerebrosidase in cultured blastoid cells (10-30 nmol substrate hydrolyzed/h.mg protein) resembles closely values observed for
-
Campbell, Jessica K; Rogers, Randy B; Lila, Mary Ann; Erdman, John W
2006-02-08
This work describes the development and utilization of a plant cell culture production approach to biosynthesize and radiolabel phytoene and phytofluene for prostate cancer cell culture studies. The herbicide norflurazon was added to established cell suspension cultures of tomato (Lycopersicon esculentum cv. VFNT cherry), to induce the biosynthesis and accumulation of the lycopene precursors, phytoene and phytofluene, in their natural isomeric forms (15-cis-phytoene and two cis-phytofluene isomers). Norflurazon concentrations, solvent carrier type and concentration, and duration of culture exposure to norflurazon were screened to optimize phytoene and phytofluene synthesis. Maximum yields of both phytoene and phytofluene were achieved after 7 days of treatment with 0.03 mg norflurazon/40 mL fresh medium, provided in 0.07% solvent carrier. Introduction of 14C-sucrose to the tomato cell culture medium enabled the production of 14C-labeled phytoene for subsequent prostate tumor cell uptake studies. In DU 145 prostate tumor cells, it was determined that 15-cis-phytoene and an oxidized product of phytoene were taken up and partially metabolized by the cells. The ability to biosynthesize, radiolabel, and isolate these carotenoids from tomato cell cultures is a novel, valuable methodology for further in vitro and in vivo investigations into the roles of phytoene and phytofluene in cancer chemoprevention.
-
Advantages and challenges of microfluidic cell culture in polydimethylsiloxane devices.
Halldorsson, Skarphedinn; Lucumi, Edinson; Gómez-Sjöberg, Rafael; Fleming, Ronan M T
2015-01-15
Culture of cells using various microfluidic devices is becoming more common within experimental cell biology. At the same time, a technological radiation of microfluidic cell culture device designs is currently in progress. Ultimately, the utility of microfluidic cell culture will be determined by its capacity to permit new insights into cellular function. Especially insights that would otherwise be difficult or impossible to obtain with macroscopic cell culture in traditional polystyrene dishes, flasks or well-plates. Many decades of heuristic optimization have gone into perfecting conventional cell culture devices and protocols. In comparison, even for the most commonly used microfluidic cell culture devices, such as those fabricated from polydimethylsiloxane (PDMS), collective understanding of the differences in cellular behavior between microfluidic and macroscopic culture is still developing. Moving in vitro culture from macroscopic culture to PDMS based devices can come with unforeseen challenges. Changes in device material, surface coating, cell number per unit surface area or per unit media volume may all affect the outcome of otherwise standard protocols. In this review, we outline some of the advantages and challenges that may accompany a transition from macroscopic to microfluidic cell culture. We focus on decisive factors that distinguish macroscopic from microfluidic cell culture to encourage a reconsideration of how macroscopic cell culture principles might apply to microfluidic cell culture. Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.
-
The Culture of Translational Science Research: Participants' Stories.
Kotarba, Joseph A; Wooten, Kevin; Freeman, Jean; Brasier, Allan R
2013-01-01
We apply a symbolic interactionist framework and a qualitative methodology to the examination of the everyday reality of translational science research (TSR). This is a growing scientific movement that aims to facilitate the efficient application of basic research to clinical service design and delivery. We describe the emerging culture of translational research at a mid-size medical center that received a Clinical and Translational Science Award from the National Institutes of Health. The stories related by scientists, clinicians, and students in interviews indicate that they make sense of the emerging inter- and cross-disciplinary, team-oriented culture of TSR through the refinement and redefinition of the significant symbols that inform their work while they attempt to master translational research by addressing the dilemmas it produces for them and their work. We see the strength, currency, adaptability, and energy of the core self-definition of "scientist" to be significant in shaping the emerging culture of translational research. We conclude by celebrating the value of interpretive ethnography for evaluation research.
-
Isolation and culture of larval cells from C. elegans.
Directory of Open Access Journals (Sweden)
Sihui Zhang
Full Text Available Cell culture is an essential tool to study cell function. In C. elegans the ability to isolate and culture cells has been limited to embryonically derived cells. However, cells or blastomeres isolated from mixed stage embryos terminally differentiate within 24 hours of culture, thus precluding post-embryonic stage cell culture. We have developed an efficient and technically simple method for large-scale isolation and primary culture of larval-stage cells. We have optimized the treatment to maximize cell number and minimize cell death for each of the four larval stages. We obtained up to 7.8×10(4 cells per microliter of packed larvae, and up to 97% of adherent cells isolated by this method were viable for at least 16 hours. Cultured larval cells showed stage-specific increases in both cell size and multinuclearity and expressed lineage- and cell type-specific reporters. The majority (81% of larval cells isolated by our method were muscle cells that exhibited stage-specific phenotypes. L1 muscle cells developed 1 to 2 wide cytoplasmic processes, while L4 muscle cells developed 4 to 14 processes of various thicknesses. L4 muscle cells developed bands of myosin heavy chain A thick filaments at the cell center and spontaneously contracted ex vivo. Neurons constituted less than 10% of the isolated cells and the majority of neurons developed one or more long, microtubule-rich protrusions that terminated in actin-rich growth cones. In addition to cells such as muscle and neuron that are high abundance in vivo, we were also able to isolate M-lineage cells that constitute less than 0.2% of cells in vivo. Our novel method of cell isolation extends C. elegans cell culture to larval developmental stages, and allows use of the wealth of cell culture tools, such as cell sorting, electrophysiology, co-culture, and high-resolution imaging of subcellular dynamics, in investigation of post-embryonic development and physiology.
-
Relevance of Piagetian cross-cultural psychology to the humanities and social sciences.
Oesterdiekhoff, Georg W
2013-01-01
Jean Piaget held views according to which there are parallels between ontogeny and the historical development of culture, sciences, and reason. His books are full of remarks and considerations about these parallels, with reference to many logical, physical, social, and moral phenomena.This article explains that Piagetian cross-cultural psychology has delivered the decisive data needed to extend the research interests of Piaget. These data provide a basis for reconstructing not only the history of sciences but also the history of religion, politics, morals, culture, philosophy, and social change and the emergence of industrial society. Thus, it is possible to develop Piagetian theory as a historical anthropology in order to provide a basis for the humanities and social sciences.
-
Cognitive science and the cultural nature of music.
Cross, Ian
2012-10-01
The vast majority of experimental studies of music to date have explored music in terms of the processes involved in the perception and cognition of complex sonic patterns that can elicit emotion. This paper argues that this conception of music is at odds both with recent Western musical scholarship and with ethnomusicological models, and that it presents a partial and culture-specific representation of what may be a generic human capacity. It argues that the cognitive sciences must actively engage with the problems of exploring music as manifested and conceived in the broad spectrum of world cultures, not only to elucidate the diversity of music in mind but also to identify potential commonalities that could illuminate the relationships between music and other domains of thought and behavior. Copyright © 2012 Cognitive Science Society, Inc.
-
Cinema and science, nature and culture
Directory of Open Access Journals (Sweden)
Marcio Barreto
2017-05-01
Full Text Available http://dx.doi.org/10.5007/1807-1384.2017v14n2p19 Faced with contemporary issues that call into question the boundaries between nature and culture, man and machine, society and environment, cinema is taken here as a guide for reflection on modern science by Bergson and Deleuze’s texts, among other authors. Operating the passage from science to art and from art to science, cinema is here considered as a technical object and technical object as instrument to magnify perception. To highlight the artificiality of these borders, Newton's gravitational law is presented in its physical and metaphysical implications by the writings of Betty Dobbs and Newton himself. Through the lens of cinema, especially on movies such as 2001 A Space Odyssey, by Stanley Kubrick and Out of the present, by Andrei Ujica, the article shows that the power of cinema contributes to the perception that dimensions of science, which go beyond their alliances with the State and the market, are relevant to the challenges on the horizon of the 21’s century.
-
The four cultures: Public engagement with science only, art only, neither, or both museums.
Shein, Paichi Pat; Li, Yuh-Yuh; Huang, Tai-Chu
2015-11-01
This study uses an art-and-science comparative lens to understand the science culture, particularly the public engagement with science museums. A representational Taiwanese sample of 1863 subjects was categorized into "four cultures," who visit science only, art only, neither, or both museums, resulting in six multivariate logistic regression models. Knowledge of science, interests in scientific and social issues, and socio-demographic variables were considered in the models. Adults with children and males prefer science museums, females prefer art museums, and the young and urban intellects show no strong preference, appearing to be open to both science and art museums. The findings show the complex decisions the public make in visiting museums. It is no longer a strictly science or art decision, as framed by Snow's "The Two Cultures" argument; rather, the possibility of visiting both museums has emerged, a phenomenon we describe as cognitive polyphasia. © The Author(s) 2015.
-
Bowey-Dellinger, Kristen; Dixon, Luke; Ackerman, Kristin; Vigueira, Cynthia; Suh, Yewseok K; Lyda, Todd; Sapp, Kelli; Grider, Michael; Crater, Dinene; Russell, Travis; Elias, Michael; Coffield, V McNeil; Segarra, Verónica A
2017-01-01
Undergraduate students learn about mammalian cell culture applications in introductory biology courses. However, laboratory modules are rarely designed to provide hands-on experience with mammalian cells or teach cell culture techniques, such as trypsinization and cell counting. Students are more likely to learn about cell culture using bacteria or yeast, as they are typically easier to grow, culture, and manipulate given the equipment, tools, and environment of most undergraduate biology laboratories. In contrast, the utilization of mammalian cells requires a dedicated biological safety cabinet and rigorous antiseptic techniques. For this reason, we have devised a laboratory module and method herein that familiarizes students with common cell culture procedures, without the use of a sterile hood or large cell culture facility. Students design and perform a time-efficient inquiry-based cell viability experiment using HeLa cells and tools that are readily available in an undergraduate biology laboratory. Students will become familiar with common techniques such as trypsinizing cells, cell counting with a hemocytometer, performing serial dilutions, and determining cell viability using trypan blue dye. Additionally, students will work with graphing software to analyze their data and think critically about the mechanism of death on a cellular level. Two different adaptations of this inquiry-based lab are presented-one for non-biology majors and one for biology majors. Overall, these laboratories aim to expose students to mammalian cell culture and basic techniques and help them to conceptualize their application in scientific research.
-
Directory of Open Access Journals (Sweden)
Erna Bastian
1971-06-01
Full Text Available Examina-se o conceito do relativismo cultural sob bases filosóficas e sua validade nas ciências da saúde. Analisam-se os têrmos absoluto, relativo e universal, chegando-se a um modêlo operacional do relativismo cultural nas ciências da saúde.The concept of cultural relativism on a philosophical basis and its validity in health sciences, is examined. The concepts of the absolute, the relative and the universal are analysed and a working model for cultural relativism in the health sciences is attained.
-
Multizone Paper Platform for 3D Cell Cultures
Derda, Ratmir; Hong, Estrella; Mwangi, Martin; Mammoto, Akiko; Ingber, Donald E.; Whitesides, George M.
2011-01-01
In vitro 3D culture is an important model for tissues in vivo. Cells in different locations of 3D tissues are physiologically different, because they are exposed to different concentrations of oxygen, nutrients, and signaling molecules, and to other environmental factors (temperature, mechanical stress, etc). The majority of high-throughput assays based on 3D cultures, however, can only detect the average behavior of cells in the whole 3D construct. Isolation of cells from specific regions of 3D cultures is possible, but relies on low-throughput techniques such as tissue sectioning and micromanipulation. Based on a procedure reported previously (“cells-in-gels-in-paper” or CiGiP), this paper describes a simple method for culture of arrays of thin planar sections of tissues, either alone or stacked to create more complex 3D tissue structures. This procedure starts with sheets of paper patterned with hydrophobic regions that form 96 hydrophilic zones. Serial spotting of cells suspended in extracellular matrix (ECM) gel onto the patterned paper creates an array of 200 micron-thick slabs of ECM gel (supported mechanically by cellulose fibers) containing cells. Stacking the sheets with zones aligned on top of one another assembles 96 3D multilayer constructs. De-stacking the layers of the 3D culture, by peeling apart the sheets of paper, “sections” all 96 cultures at once. It is, thus, simple to isolate 200-micron-thick cell-containing slabs from each 3D culture in the 96-zone array. Because the 3D cultures are assembled from multiple layers, the number of cells plated initially in each layer determines the spatial distribution of cells in the stacked 3D cultures. This capability made it possible to compare the growth of 3D tumor models of different spatial composition, and to examine the migration of cells in these structures. PMID:21573103
-
Well-ordered science and Indian epistemic cultures: toward a polycentered history of science.
Ganeri, Jonardon
2013-06-01
This essay defends the view that "modern science," as with modernity in general, is a polycentered phenomenon, something that appears in different forms at different times and places. It begins with two ideas about the nature of rational scientific inquiry: Karin Knorr Cetina's idea of "epistemic cultures," and Philip Kitcher's idea of science as "a system of public knowledge," such knowledge as would be deemed worthwhile by an ideal conversation among the whole public under conditions of mutual engagement. This account of the nature of scientific practice provides us with a new perspective from which to understand key elements in the philosophical project of Jaina logicians in the seventh, eighth, and ninth centuries C.E. Jaina theory seems exceptionally well targeted onto two of the key constituents in the ideal conversation--the classification of all human points of view and the representation of end states of the deliberative process. The Buddhist theory of the Kathāvatthu contributes to Indian epistemic culture in a different way: by supplying a detailed theory of how human dialogical standpoints can be revised in the ideal conversation, an account of the phenomenon Kitcher labels "tutoring." Thus science in India has its own history, one that should be studied in comparison and contrast with the history of science in Europe. In answer to Joseph Needham, it was not 'modern science' which failed to develop in India or China but rather non-well-ordered science, science as unconstrained by social value and democratic consent. What I argue is that this is not a deficit in the civilisational histories of these countries, but a virtue.
-
Krüger-Genge, A; Fuhrmann, R; Jung, F; Franke, R P
2015-01-01
The evaluation of the interaction of human, venous endothelial cells (HUVEC) with body foreign materials on the cellular level cannot be performed in vivo, but is investigated in vitro under standard culture conditions. To maintain the vitality, proliferation and morphology of HUVEC seeded on body foreign substrates over days, the cell culture medium is usually exchanged every second day. It is well known, that alterations in the microenvironment of cells bear the risk of influencing cell morphology and function. In the current study the influence of cell culture medium exchange on HUVEC cytoskeletal microfilament structure and function was investigated. HUVEC in the third passage were seeded on extracellular matrix (ECM) - which was secreted from bovine corneal endothelial cells on glass- until functional confluence was reached. The experiment started 11 days after HUVEC seeding with an exchange of the cell culture medium followed by a staining of the actin microfilaments with phalloidin-rhodamin 1.5 and 5 minutes after medium exchange. The microfilaments were documented by use of an Olympus microscope (IMT-2) equipped with a UV lamp and online connected to a TV chain (Sony XC 50 ST/monochrome) implying an OPTIMAS - Image analysis system. Prostacyclin was analysed in the cell culture supernatant. 1.5 min after culture medium exchange in the functionally confluent cultures a slight disturbance of the actin microfilament structure with a broadening of the marginal filament band, a partial disconnection of cell-cell contacts and the appearance of intercellular fenestrations were observed. 5 minutes after medium exchange a redevelopment of the slightly disturbed microfilament structure with a condensation and narrowing of the marginal filament band was seen. 12 h later a further consolidation of the microfilament structure occurred. In addition, a perturbation of the cultured HUVEC occurred after cell culture medium exchange. The prostacyclin concentration in the
-
Jackson, Matthew C.; Galvez, Gino; Landa, Isidro; Buonora, Paul; Thoman, Dustin B.
2016-01-01
Recent research suggests that underrepresented minority (URM) college students, and especially first-generation URMs, may lose motivation to persist if they see science careers as unable to fulfill culturally relevant career goals. In the present study, we used a mixed-methods approach to explore patterns of motivation to pursue physical and life sciences across ethnic groups of freshman college students, as moderated by generational status. Results from a longitudinal survey (N = 249) demonstrated that freshman URM students who enter with a greater belief that science can be used to help their communities identified as scientists more strongly over time, but only among first-generation college students. Analysis of the survey data were consistent with content analysis of 11 transcripts from simultaneously conducted focus groups (N = 67); together, these studies reveal important differences in motivational characteristics both across and within ethnicity across educational generation status. First-generation URM students held the strongest prosocial values for pursuing a science major (e.g., giving back to the community). URM students broadly reported additional motivation to increase the status of their family (e.g., fulfilling aspirations for a better life). These findings demonstrate the importance of culturally connected career motives and for examining intersectional identities to understand science education choices and inform efforts to broaden participation. PMID:27543631
-
Malama I Ka `Aina: Fostering the Culture-Science connection
Bruno, B.; Chinn, P.
2005-12-01
The Malama I Ka `Aina Project (Caring for the land, or sustainability) aims to improve and expand the education of Hawai`i's children by developing and disseminating standards-based, culturally relevant science curricular materials based on an understanding and appreciation of the ways in which traditional Hawaiians interacted with their environment for sustainability. Key concepts include the role of water and the ahupua`a (traditional Hawaiian system of land management), and a culture-based sense of place that includes knowledge of and connection to the land. Elementary, middle, high school and University of Hawai`i teachers work together to develop and implement curricula that are especially relevant to a particular school's science program and issues, e.g., invasive species, students, community and/or geographical location. Participants (typically a mix of teachers, education majors and science majors) enroll in Malama I Ka `Aina, a three-credit course offered through the University of Hawai`i`s Dept. of Curriculum Studies and applicable toward a Bachelor's or Master's degree. This course (team taught by scientists, cultural experts and educational professionals) enables participants to: (1) Study Hawai`i`s unique geology, geography and environmental issues in the context of Hawaiian culture and post Western contact; (2) Use course knowledge to develop, teach and assess Hawaii-oriented, project-based, inquiry activities that address the Hawaii Science Content Standards; (3) Gain an appreciation for the scientific method, and the curiosity that drives science (4) Use educational technology such as PowerPoint, graphing packages and web authoring software to develop electronic resources for educational activities. A sample of the lessons developed by course participants can be found on http://malama.hawaii.edu/schools/index2.html. This project is based at the University of Hawai`i College of Education and funded by an award to P. Chinn by the US Department of
-
Mutation in cultured mammalian cells
International Nuclear Information System (INIS)
Nakamura, N.; Okada, S.
1982-01-01
Mammalian cell cultures were exposed to gamma-rays at various dose rates. Dose-rate effects were observed in cultured somatic cells of the mouse for cell killing and mutations resistant to 6-thioguanine (TGsup(r)) and to methotrexate (MTXsup(r)). Linear quadratic model may be applied to cell killing and TGsup(r) mutations in some cases but can not explain the whole data. Results at low doses with far low dose-rate were not predictable from data at high doses with acute or chronic irradiation. Radioprotective effects of dimethyl sulfoxide were seen only after acute exposure but not after chronic one, suggesting that damages by indirect action of radiations may be potentially reparable by cells. TGsup(r) mutations seem to contain gross structural changes whereas MTXsup(r) ones may have smaller alterations. (Namekawa, K.)
-
Cell-free DNA in a three-dimensional spheroid cell culture model
DEFF Research Database (Denmark)
Aucamp, Janine; Calitz, Carlemi; Bronkhorst, Abel J.
2017-01-01
Background Investigating the biological functions of cell-free DNA (cfDNA) is limited by the interference of vast numbers of putative sources and causes of DNA release into circulation. Utilization of three-dimensional (3D) spheroid cell cultures, models with characteristics closer to the in vivo...... cultures can serve as effective, simplified in vivo-simulating “closed-circuit” models since putative sources of cfDNA are limited to only the targeted cells. In addition, cfDNA can also serve as an alternative or auxiliary marker for tracking spheroid growth, development and culture stability. Biological...... significance 3D cell cultures can be used to translate “closed-circuit” in vitro model research into data that is relevant for in vivo studies and clinical applications. In turn, the utilization of cfDNA during 3D culture research can optimize sample collection without affecting the stability of the growth...
-
Changing the Culture of Science Communication Training for Junior Scientists
Bankston, Adriana; McDowell, Gary S.
2018-01-01
Being successful in an academic environment places many demands on junior scientists. Science communication currently may not be adequately valued and rewarded, and yet communication to multiple audiences is critical for ensuring that it remains a priority in today’s society. Due to the potential for science communication to produce better scientists, facilitate scientific progress, and influence decision-making at multiple levels, training junior scientists in both effective and ethical science communication practices is imperative, and can benefit scientists regardless of their chosen career path. However, many challenges exist in addressing specific aspects of this training. Principally, science communication training and resources should be made readily available to junior scientists at institutions, and there is a need to scale up existing science communication training programs and standardize core aspects of these programs across universities, while also allowing for experimentation with training. We propose a comprehensive core training program be adopted by universities, utilizing a centralized online resource with science communication information from multiple stakeholders. In addition, the culture of science must shift toward greater acceptance of science communication as an essential part of training. For this purpose, the science communication field itself needs to be developed, researched and better understood at multiple levels. Ultimately, this may result in a larger cultural change toward acceptance of professional development activities as valuable for training scientists. PMID:29904538
-
Changing the Culture of Science Communication Training for Junior Scientists.
Bankston, Adriana; McDowell, Gary S
2018-01-01
Being successful in an academic environment places many demands on junior scientists. Science communication currently may not be adequately valued and rewarded, and yet communication to multiple audiences is critical for ensuring that it remains a priority in today's society. Due to the potential for science communication to produce better scientists, facilitate scientific progress, and influence decision-making at multiple levels, training junior scientists in both effective and ethical science communication practices is imperative, and can benefit scientists regardless of their chosen career path. However, many challenges exist in addressing specific aspects of this training. Principally, science communication training and resources should be made readily available to junior scientists at institutions, and there is a need to scale up existing science communication training programs and standardize core aspects of these programs across universities, while also allowing for experimentation with training. We propose a comprehensive core training program be adopted by universities, utilizing a centralized online resource with science communication information from multiple stakeholders. In addition, the culture of science must shift toward greater acceptance of science communication as an essential part of training. For this purpose, the science communication field itself needs to be developed, researched and better understood at multiple levels. Ultimately, this may result in a larger cultural change toward acceptance of professional development activities as valuable for training scientists.
-
MEMS-based dynamic cell-to-cell culture platforms using electrochemical surface modifications
International Nuclear Information System (INIS)
Chang, Jiyoung; Lin, Liwei; Yoon, Sang-Hee; Mofrad, Mohammad R K
2011-01-01
MEMS-based biological platforms with the capability of both spatial placements and time releases of living cells for cell-to-cell culture experiments have been designed and demonstrated utilizing electrochemical surface modification effects. The spatial placement is accomplished by electrochemical surface modification of substrate surfaces to be either adhesive or non-adhesive for living cells. The time control is achieved by the electrical activation of the selective indium tin oxide co-culture electrode to allow the migration of living cells onto the electrode to start the cell-to-cell culture studies. Prototype devices have a three-electrode design with an electrode size of 50 × 50 µm 2 and the separation gaps of 2 µm between them. An electrical voltage of −1.5 V has been used to activate the electrodes independently and sequentially to demonstrate the dynamic cell-to-cell culture experiments of NIH 3T3 fibroblast and Madin Darby canine kidney cells. As such, this MEMS platform could be a basic yet versatile tool to characterize transient cell-to-cell interactions
-
Microfluidic engineered high cell density three-dimensional neural cultures
Cullen, D. Kacy; Vukasinovic, Jelena; Glezer, Ari; La Placa, Michelle C.
2007-06-01
Three-dimensional (3D) neural cultures with cells distributed throughout a thick, bioactive protein scaffold may better represent neurobiological phenomena than planar correlates lacking matrix support. Neural cells in vivo interact within a complex, multicellular environment with tightly coupled 3D cell-cell/cell-matrix interactions; however, thick 3D neural cultures at cell densities approaching that of brain rapidly decay, presumably due to diffusion limited interstitial mass transport. To address this issue, we have developed a novel perfusion platform that utilizes forced intercellular convection to enhance mass transport. First, we demonstrated that in thick (>500 µm) 3D neural cultures supported by passive diffusion, cell densities =104 cells mm-3), continuous medium perfusion at 2.0-11.0 µL min-1 improved viability compared to non-perfused cultures (p death and matrix degradation. In perfused cultures, survival was dependent on proximity to the perfusion source at 2.00-6.25 µL min-1 (p 90% viability in both neuronal cultures and neuronal-astrocytic co-cultures. This work demonstrates the utility of forced interstitial convection in improving the survival of high cell density 3D engineered neural constructs and may aid in the development of novel tissue-engineered systems reconstituting 3D cell-cell/cell-matrix interactions.
-
Cell sources for in vitro human liver cell culture models
Freyer, Nora; Damm, Georg; Seehofer, Daniel; Knöspel, Fanny
2016-01-01
In vitro liver cell culture models are gaining increasing importance in pharmacological and toxicological research. The source of cells used is critical for the relevance and the predictive value of such models. Primary human hepatocytes (PHH) are currently considered to be the gold standard for hepatic in vitro culture models, since they directly reflect the specific metabolism and functionality of the human liver; however, the scarcity and difficult logistics of PHH have driven researchers to explore alternative cell sources, including liver cell lines and pluripotent stem cells. Liver cell lines generated from hepatomas or by genetic manipulation are widely used due to their good availability, but they are generally altered in certain metabolic functions. For the past few years, adult and pluripotent stem cells have been attracting increasing attention, due their ability to proliferate and to differentiate into hepatocyte-like cells in vitro. However, controlling the differentiation of these cells is still a challenge. This review gives an overview of the major human cell sources under investigation for in vitro liver cell culture models, including primary human liver cells, liver cell lines, and stem cells. The promises and challenges of different cell types are discussed with a focus on the complex 2D and 3D culture approaches under investigation for improving liver cell functionality in vitro. Finally, the specific application options of individual cell sources in pharmacological research or disease modeling are described. PMID:27385595
-
Growth of melanocytes in human epidermal cell cultures
International Nuclear Information System (INIS)
Staiano-Coico, L.; Hefton, J.M.; Amadeo, C.; Pagan-Charry, I.; Madden, M.R.; Cardon-Cardo, C.
1990-01-01
Epidermal cell cultures were grown in keratinocyte-conditioned medium for use as burn wound grafts; the melanocyte composition of the grafts was studied under a variety of conditions. Melanocytes were identified by immunohistochemistry based on a monoclonal antibody (MEL-5) that has previously been shown to react specifically with melanocytes. During the first 7 days of growth in primary culture, the total number of melanocytes in the epidermal cultures decreased to 10% of the number present in normal skin. Beginning on day 2 of culture, bipolar melanocytes were present at a mean cell density of 116 +/- 2/mm2; the keratinocyte to melanocyte ratio was preserved during further primary culture and through three subpassages. Moreover, exposure of cultures to mild UVB irradiation stimulated the melanocytes to proliferate, suggesting that the melanocytes growing in culture maintained their responsiveness to external stimuli. When the sheets of cultured cells were enzymatically detached from the plastic culture flasks before grafting, melanocytes remained in the basal layer of cells as part of the graft applied to the patient
-
Cell Culture as an Alternative in Education.
Nardone, Roland M.
1990-01-01
Programs that are intended to inform and provide "hands-on" experience for students and to facilitate the introduction of cell culture-based laboratory exercises into the high school and college laboratory are examined. The components of the CellServ Program and the Cell Culture Toxicology Training Programs are described. (KR)
-
Science cafés. Cross-cultural adaptation and educational applications
Directory of Open Access Journals (Sweden)
M. Norton
2009-10-01
Full Text Available Tokyo Institute of Technology (TokyoTech has been developing a number of methodologies to teach graduate students the theory and practice of science communication since 2005. One of the tools used is the science café, where students are taught about the background based primarily on theoretical models developed in the UK. They then apply that knowledge and adapt it the Japanese cultural context and plan, execute and review outcomes as part of their course. In this paper we review 4 years of experience in using science cafés in this educational context; we review the background to the students’ decision-making and consensus-building process towards deciding on the style and subject to be used, and the value this has in illuminating the cultural influences on the science café design and implementation. We also review the value of the science café as an educational tool and conclude that it has contributed to a number of teaching goals related to both knowledge and the personal skills required to function effectively in an international environment.
-
International Nuclear Information System (INIS)
Fujiwara, Daisuke; Kato, Kazunori; Nohara, Shigeo; Iwanuma, Yoshimi; Kajiyama, Yoshiaki
2013-01-01
Highlights: •Spheroids were created from esophageal carcinoma cells using NanoCulture® Plates. •The proportion of strongly ALDH-positive cells increased in 3-D culture. •Expression of cancer stem cell-related genes was enhanced in 3-D culture. •CA-9 expression was enhanced, suggesting hypoxia had been induced in 3-D culture. •Drug resistance was increased. 3-D culture is useful for inducing cancer stem cells. -- Abstract: In recent years, research on resistance to chemotherapy and radiotherapy in cancer treatment has come under the spotlight, and researchers have also begun investigating the relationship between resistance and cancer stem cells. Cancer stem cells are assumed to be present in esophageal cancer, but experimental methods for identification and culture of these cells have not yet been established. To solve this problem, we created spheroids using a NanoCulture® Plate (NCP) for 3-dimensional (3-D) cell culture, which was designed as a means for experimentally reproducing the 3-D structures found in the body. We investigated the potential for induction of cancer stem cells from esophageal cancer cells. Using flow cytometry we analyzed the expression of surface antigen markers CD44, CD133, CD338 (ABCG2), CD318 (CDCP1), and CD326 (EpCAM), which are known cancer stem cell markers. None of these surface antigen markers showed enhanced expression in 3-D cultured cells. We then analyzed aldehyde dehydrogenase (ALDH) enzymatic activity using the ALDEFLUOR reagent, which can identify immature cells such as stem cells and precursor cells. 3-D-cultured cells were strongly positive for ALDH enzyme activity. We also analyzed the expression of the stem cell-related genes Sox-2, Nanog, Oct3/4, and Lin28 using RT-PCR. Expression of Sox-2, Nanog, and Lin28 was enhanced. Analysis of expression of the hypoxic surface antigen marker carbonic anhydrase-9 (CA-9), which is an indicator of cancer stem cell induction and maintenance, revealed that CA-9 expression
-
Science, culture and the search for life on other worlds
Traphagan, John W
2016-01-01
This book explores humanity’s thoughts and ideas about extraterrestrial life, paying close attention to the ways science and culture interact with one another to create a context of imagination and discovery related to life on other worlds. Despite the recent explosion in our knowledge of other planets and the seeming era of discovery in which we live, to date we have found no concrete evidence that we are not alone. Our thinking about life on other worlds has been and remains the product of a combination of scientific investigation and human imagination shaped by cultural values--particularly values of exploration and discovery connected to American society. The rapid growth in our awareness of other worlds makes this a crucial moment to think about and assess the influence of cultural values on the scientific search for extraterrestrial life. Here the author considers the junction of science and culture with a focus on two main themes: (1) the underlying assumptions, many of which are tacitly based upon c...
-
How to compare the social foundations of science culture: A trial with five cities in Korea.
Song, Jinwoong; Chung, Minkyung; Choi, Eunjeong; Kim, Leekyoung; Cho, Sook-Kyoung
2013-01-01
Though there have been several indicator systems to monitor the status quo of science and technology and of scientific literacy, few are especially designed for science culture, especially for its social dimension. Furthermore there is little agreement on how to measure it. In a previous study, an indicator system, SCI (Science Culture Indicators), had been developed to monitor the status quo of the science culture of a nation at both individual and social dimensions. The purpose of this study was to explore a practical way to measure and compare local cities' social foundation of science culture by revising and standardizing the social dimension of SCI and by applying it to five metropolitan cities in Korea. Despite some limits, the results of this study appear not only to reflect the cities' current situations but also to show the strength and weakness of their social foundation of science culture.
-
Charlton, Bruce G
2006-01-01
Educational expansion in western countries has been achieved mainly by adding years to full-time education; however, this process has probably reduced efficiency. Sooner or later, efficiency must improve, with a greater educational attainment per year. Future societies will probably wish more people to study science throughout high school (aged c. 11-19 years) and the first college degree. 'Science' may be defined as any abstract, systematic and research-based discipline: including mathematics, statistics and the natural sciences, economics, music theory, linguistics, and the conceptual or quantitative social sciences. Since formal teaching is usually necessary to learn science, science education should be regarded as the core function of high schools. One standard way to improve efficiency is the 'division of labour', with increased specialization of function. Modern schools are already specialized: teachers are specialized according to age-group taught, subject matter expertise, and administrative responsibilities. School students are stratified by age and academic aptitude. I propose a further institutional division of school function between science education, and cultural education (including education in arts, sports, ethics, social interaction and good citizenship). Existing schools might split into 'science school' and 'culture school', reflected in distinct buildings and zones, separate administrative structures, and the recruitment of differently-specialized teaching personnel. Science school would be distinguished by its focus on education in disciplines which promote abstract systematic cognition. All students would spend some part of each day (how much would depend on their aptitude and motivation) in the 'science school'; experiencing a traditional-style, didactic, disciplined and rigorous academic education. The remainder of the students' time at school would be spent in the cultural division, which would focus on broader aspects, and aim to generate
-
PECULIARITIES OF SECONDARY METABOLITES BIOSYNTHESIS IN PLANT CELL CULTURES
Directory of Open Access Journals (Sweden)
A.M. NOSOV
2014-06-01
Full Text Available metabolites formation in plant cell cultures of Panax spp., (ginsenosides; Dioscorea deltoidea (steroid glycosides; Ajuga reptans, Serratula coronata, Rhaponticum carthamoides (ecdisteroids; Polyscias spp., (triterpene glycosides, Taxus spp. (taxoids, Stevia rebaudiana (diterpene steviol-glycosides, Stephania glabra (alkaloids. They are some regular trends of secondary metabolites synthesis in the plant cell culture:It can be noted the stable synthesis of the compound promoting cell proliferation. Indeed, cell cultures of Dioscorea deltoidea were demonstrated to accumulate only furostanol glycosides, which promoted cell division. Furostanol glycoside content of Dioscorea strain DM-0.5 was up to 6 - 12% by dry biomass.Panax ginseng and P. japonicus plant cell cultures synthesize as minimum seven triterpene glycosides (ginsenosides, the productivity of these compounds was up to 6.0 - 8.0% on dry biomass.By contrast, the detectable synthesis of diterpene steviol-glycosides in cultivated cells of Stevia rebaudiana initiated in the mixotrophic cultures during chloroplast formation only.Despite these differences, or mainly due to them, plant cell cultures have become an attractive source of phytochemicals in alternative to collecting wild plants. It provides a guideline to bioreactor-based production of isoprenoids using undifferentiated plant cell cultures.
-
Substrate utilisation by plant-cell cultures
Energy Technology Data Exchange (ETDEWEB)
Fowler, M W
1982-01-01
Plant cell cultures have been grown on a wide range of carbon sources in addition to the traditional ones of sucrose and glucose. Biomass yields and growth rates vary greatly between the different carbon sources and there is a variation in response between different cell cultures to individual carbon sources. Some attempts have been made to grow cell cultures on 'waste' and related carbon sources, such as lactose, maltose, starch, molasses and milk whey. Only maltose was found to support growth to anything near the levels observed with glucose and sucrose. In the case of molasses carbon source cell growth was either non-existent or only just measurable. All the data point to glucose as being the most suitable carbon source, principally on the grounds of biomass yield and growth rate. It should be noted, however, that other carbon sources do appear to have a major (positive) influence on natural product synthesis. Uptake into the cell is an important aspect of carbohydrate utilisation. There is strong evidence that from disaccharides upwards, major degradation to smaller units occurs before uptake. In some cases the necessary enzymes appear to be excreted into the culture broth, in others they may be located within the cell wall; invertase that hydrolyses sucrose is a good example. Once the products of carbohydrate degradation and mobilisation enter the cell they may suffer one of two fates, oxidation or utilisation for biosynthesis. The precise split between these two varies depending on such factors as cell growth rate, cell size, nutrient broth composition and carbohydrate status of the cells. In general rapidly growing cells have a high rate of oxidation, whereas cells growing more slowly tend to be more directed towards biosynthesis. Carbohydrate utilisation is a key area of study, underpinning as it does both biomass yield and natural product synthesis. (Refs. 13).
-
Biona-C Cell Culture pH Monitoring System
Friedericks, C.
1999-01-01
Sensors 2000! is developing a system to demonstrate the ability to perform accurate, real-time measurements of pH and CO2 in a cell culture media in Space. The BIONA-C Cell Culture pH Monitoring System consists of S2K! developed ion selective sensors and control electronics integrated with the fluidics of a cell culture system. The integrated system comprises a "rail" in the Cell Culture Module (CCM) of WRAIR (Space Biosciences of Walter Read Army Institute of Research). The CCM is a Space Shuttle mid-deck locker experiment payload. The BIONA-C is displayed along with associated graphics and text explanations. The presentation will stimulate interest in development of sensor technology for real-time cell culture measurements. The transfer of this technology to other applications will also be of interest. Additional information is contained in the original document.
-
Predictors of cultural capital on science academic achievement at the 8th grade level
Misner, Johnathan Scott
The purpose of the study was to determine if students' cultural capital is a significant predictor of 8th grade science achievement test scores in urban locales. Cultural capital refers to the knowledge used and gained by the dominant class, which allows social and economic mobility. Cultural capital variables include magazines at home and parental education level. Other variables analyzed include socioeconomic status (SES), gender, and English language learners (ELL). This non-experimental study analyzed the results of the 2011 Eighth Grade Science National Assessment of Educational Progress (NAEP). The researcher analyzed the data using a multivariate stepwise regression analysis. The researcher concluded that the addition of cultural capital factors significantly increased the predictive power of the model where magazines in home, gender, student classified as ELL, parental education level, and SES were the independent variables and science achievement was the dependent variable. For alpha=0.05, the overall test for the model produced a R2 value of 0.232; therefore the model predicted 23.2% of variance in science achievement results. Other major findings include: higher measures of home resources predicted higher 2011 NAEP eighth grade science achievement; males were predicted to have higher 2011 NAEP 8 th grade science achievement; classified ELL students were predicted to score lower on the NAEP eight grade science achievement; higher parent education predicted higher NAEP eighth grade science achievement; lower measures of SES predicted lower 2011 NAEP eighth grade science achievement. This study contributed to the research in this field by identifying cultural capital factors that have been found to have statistical significance on predicting eighth grade science achievement results, which can lead to strategies to help improve science academic achievement among underserved populations.
-
Laundos, Tiago L; Silva, Joana; Assunção, Marisa; Quelhas, Pedro; Monteiro, Cátia; Oliveira, Carla; Oliveira, Maria J; Pêgo, Ana P; Amaral, Isabel F
2017-08-01
Embryonic stem (ES)-derived neural stem/progenitor cells (ES-NSPCs) constitute a promising cell source for application in cell therapies for the treatment of central nervous system disorders. In this study, a rotary orbital hydrodynamic culture system was applied to single-cell suspensions of ES-NSPCs, to obtain homogeneously-sized ES-NSPC cellular aggregates (neurospheres). Hydrodynamic culture allowed the formation of ES-NSPC neurospheres with a narrower size distribution than statically cultured neurospheres, increasing orbital speeds leading to smaller-sized neurospheres and higher neurosphere yield. Neurospheres formed under hydrodynamic conditions (72 h at 55 rpm) showed higher cell compaction and comparable percentages of viable, dead, apoptotic and proliferative cells. Further characterization of cellular aggregates provided new insights into the effect of hydrodynamic shear on ES-NSPC behaviour. Rotary neurospheres exhibited reduced protein levels of N-cadherin and β-catenin, and higher deposition of laminin (without impacting fibronectin deposition), matrix metalloproteinase-2 (MMP-2) activity and percentage of neuronal cells. In line with the increased MMP-2 activity levels found, hydrodynamically-cultured neurospheres showed higher outward migration on laminin. Moreover, when cultured in a 3D fibrin hydrogel, rotary neurospheres generated an increased percentage of neuronal cells. In conclusion, the application of a constant orbital speed to single-cell suspensions of ES-NSPCs, besides allowing the formation of homogeneously-sized neurospheres, promoted ES-NSPC differentiation and outward migration, possibly by influencing the expression of cell-cell adhesion molecules and the secretion of proteases/extracellular matrix proteins. These findings are important when establishing the culture conditions needed to obtain uniformly-sized ES-NSPC aggregates, either for use in regenerative therapies or in in vitro platforms for biomaterial development or
-
Protein biosynthesis in cultured human hair follicle cells.
Weterings, P J; Vermorken, A J; Bloemendal, H
1980-10-31
A new technique has been used for culturing human keratinocytes. The cells grow on the basement membrane-like capsules of bovine lenses. Lens cells were removed from the capsules by rigid trypsinization. In order to exclude any contamination with remaining living cells the isolated capsules were irradiated with X-rays at a dose of 10,000 rad. In this way human epithelial cells can be brought in culture from individual hair follicles. Since feeder cells are not used in this culture technique, the biosynthesis of keratinocyte proteins can be studied in these cultures. The newly synthesized proteins can be separated into a water-soluble, a urea-soluble, and a urea-insoluble fraction. Product analysis has been performed on the first two fractions revealing protein patterns identical to those of intact hair follicles. Product analysis of the urea-soluble fractions of microdissected hair follicles shows that the protein pattern of the cultured keratinocytes resembles the protein pattern of the hair follicle sheath. Studies on the metabolism of benzo(a)pyrene revealed that the enzyme aryl hydrocarbon hydroxylase (AHH) is present in cultured hair follicle cells. A possible use of our culture system for eventual detection of inherited predisposition for smoking-dependent lung cancer is discussed.
-
21 CFR 864.2280 - Cultured animal and human cells.
2010-04-01
... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Cultured animal and human cells. 864.2280 Section... Cultured animal and human cells. (a) Identification. Cultured animal and human cells are in vitro cultivated cell lines from the tissue of humans or other animals which are used in various diagnostic...
-
International Nuclear Information System (INIS)
Otsuka, Hidenori; Sasaki, Kohei; Okimura, Saya; Nagamura, Masako; Nakasone, Yuichi
2013-01-01
Microfabrication and micropatterning techniques in tissue engineering offer great potential for creating and controlling cellular microenvironments including cell–matrix interactions, soluble stimuli and cell–cell interactions. Here, we present a novel approach to generate layered patterning of hepatocyte spheroids on micropatterned non-parenchymal feeder cells using microfabricated poly(ethylene glycol) (PEG) hydrogels. Micropatterned PEG-hydrogel-treated substrates with two-dimensional arrays of gelatin circular domains (ϕ = 100 μm) were prepared by photolithographic method. Only on the critical structure of PEG hydrogel with perfect protein rejection, hepatocytes were co-cultured with non-parenchymal cells to be led to enhanced hepatocyte functions. Then, we investigated the mechanism of the functional enhancement in co-culture with respect to the contributions of soluble factors and direct cell–cell interactions. In particular, to elucidate the influence of soluble factors on hepatocyte function, hepatocyte spheroids underlaid with fibroblasts (NIH/3T3 mouse fibroblasts) or endothelial cells (BAECs: bovine aortic endothelial cells) were compared with physically separated co-culture of hepatocyte monospheroids with NIH3T3 or BAEC using trans-well culture systems. Our results suggested that direct heterotypic cell-to-cell contact and soluble factors, both of these between hepatocytes and fibroblasts, significantly enhanced hepatocyte functions. In contrast, direct heterotypic cell-to-cell contact between hepatocytes and endothelial cells only contributed to enhance hepatocyte functions. This patterning technique can be a useful experimental tool for applications in basic science, drug screening and tissue engineering, as well as in the design of artificial liver devices. (paper)
-
Díez, José M; Bauman, Ewa; Gajardo, Rodrigo; Jorquera, Juan I
2015-03-13
Fetal bovine serum (FBS) is an animal product used as a medium supplement. The animal origin of FBS is a concern if cultured stem cells are to be utilized for human cell therapy. Therefore, a substitute for FBS is desirable. In this study, an industrial, xeno-free, pharmaceutical-grade supplement for cell culture (SCC) under development at Grifols was tested for growth of human mesenchymal stem cells (hMSCs), cell characterization, and differentiation capacity. SCC is a freeze-dried product obtained through cold-ethanol fractionation of industrial human plasma pools from healthy donors. Bone marrow-derived hMSC cell lines were obtained from two commercial suppliers. Cell growth was evaluated by culturing hMSCs with commercial media or media supplemented with SCC or FBS. Cell viability and cell yield were assessed with an automated cell counter. Cell surface markers were studied by indirect immunofluorescence assay. Cells were cultured then differentiated into adipocytes, chondrocytes, osteoblasts, and neurons, as assessed by specific staining and microscopy observation. SCC supported the growth of commercial hMSCs. Starting from the same number of seeded cells in two consecutive passages of culture with medium supplemented with SCC, hMSC yield and cell population doubling time were equivalent to the values obtained with the commercial medium and was consistent among lots. The viability of hMSCs was higher than 90%, while maintaining the characteristic phenotype of undifferentiated hMSCs (positive for CD29, CD44, CD90, CD105, CD146, CD166 and Stro-1; negative for CD14 and CD19). Cultured hMSCs maintained the potential for differentiation into adipocytes, chondrocytes, osteoblasts, and neurons. The tested human plasma-derived SCC sustains the adequate growth of hMSCs, while preserving their differentiation capacity. SCC can be a potential candidate for cell culture supplement in advanced cell therapies.
-
Differential marker expression by cultures rich in mesenchymal stem cells
2013-01-01
Background Mesenchymal stem cells have properties that make them amenable to therapeutic use. However, the acceptance of mesenchymal stem cells in clinical practice requires standardized techniques for their specific isolation. To date, there are no conclusive marker (s) for the exclusive isolation of mesenchymal stem cells. Our aim was to identify markers differentially expressed between mesenchymal stem cell and non-stem cell mesenchymal cell cultures. We compared and contrasted the phenotype of tissue cultures in which mesenchymal stem cells are rich and rare. By initially assessing mesenchymal stem cell differentiation, we established that bone marrow and breast adipose cultures are rich in mesenchymal stem cells while, in our hands, foreskin fibroblast and olfactory tissue cultures contain rare mesenchymal stem cells. In particular, olfactory tissue cells represent non-stem cell mesenchymal cells. Subsequently, the phenotype of the tissue cultures were thoroughly assessed using immuno-fluorescence, flow-cytometry, proteomics, antibody arrays and qPCR. Results Our analysis revealed that all tissue cultures, regardless of differentiation potential, demonstrated remarkably similar phenotypes. Importantly, it was also observed that common mesenchymal stem cell markers, and fibroblast-associated markers, do not discriminate between mesenchymal stem cell and non-stem cell mesenchymal cell cultures. Examination and comparison of the phenotypes of mesenchymal stem cell and non-stem cell mesenchymal cell cultures revealed three differentially expressed markers – CD24, CD108 and CD40. Conclusion We indicate the importance of establishing differential marker expression between mesenchymal stem cells and non-stem cell mesenchymal cells in order to determine stem cell specific markers. PMID:24304471
-
Gondwe, Mzamose; Longnecker, Nancy
2015-02-01
There is no consensus in the science education research community on the meanings and representations of western science and indigenous knowledge or the relationships between them. How students interpret these relationships and their perceptions of any connections has rarely been studied. This study reports student perceptions of the meaning and relationship between scientific and cultural knowledge. Personal meaning maps adapted for small groups were conducted in seven culturally diverse schools, school years 7-9 (with students aged 12-15 years) ( n = 190), with six schools in Western Australia and one school in Malawi, Africa. Of the six Australian school groups, two comprised Australian Aboriginal students in an after-school homework programme and the other four schools had a multicultural mix of students. Students in this study identified connections between scientific and cultural knowledge and constructed connections from particular thematic areas—mainly factual content knowledge as opposed to ideas related to values, attitudes, beliefs and identity. Australian Aboriginal students made fewer connections between the two knowledge domains than Malawian students whose previous science teacher had made explicit connections in her science class. Examples from Aboriginal culture were the most dominant illustrations of cultural knowledge in Australian schools, even in school groups with students from other cultures. In light of our findings, we discuss the construction of common ground between scientific knowledge and cultural knowledge and the role of teachers as cultural brokers and travel agents. We conclude with recommendations on creating learning environments that embrace different cultural knowledges and that promote explicit and enquiring discussions of values, attitudes, beliefs and identity associated with both knowledge domains.
-
Thomay, K; Schienke, A; Vajen, B; Modlich, U; Schambach, A; Hofmann, W; Schlegelberger, B; Göhring, G
2014-01-01
The fate of cultivated primary hematopoietic stem cells (HSCs) with respect to genetic instability and telomere attrition has not yet been described in great detail. Thus, knowledge of the genetic constitution of HSCs is important when interpreting results of HSCs in culture. While establishing a cell culture model for myelodysplastic syndrome with a deletion in 5q by performing RPS14 knockdown, we found surprising data that may be of importance for any CD34+ cell culture experiments. We performed cytogenetic analyses and telomere length measurement on transduced CD34+ cells and untransduced control cells to observe the effects of long-term culturing. Initially, CD34+ cells had a normal median telomere length of about 12 kb and showed no signs of chromosomal instability. During follow-up, the median telomere length seemed to decrease and, simultaneously, increased chromosomal instability could be observed - in modified and control cells. One culture showed a clonal monosomy 7 - independent of prior RPS14 knockdown. During further culturing, it seemed that the telomeres re-elongated, and chromosomes stabilized, while TERT expression was not elevated. In summary, irrespective of our results of RPS14 knockdown in the long-term culture of CD34+ cells, it becomes clear that cell culture artefacts inducing telomere shortening and chromosomal instability have to be taken into account and regular cytogenetic analyses should always be performed. © 2013 S. Karger AG, Basel.
-
A single-cell and feeder-free culture system for monkey embryonic stem cells.
Ono, Takashi; Suzuki, Yutaka; Kato, Yosuke; Fujita, Risako; Araki, Toshihiro; Yamashita, Tomoko; Kato, Hidemasa; Torii, Ryuzo; Sato, Naoya
2014-01-01
Primate pluripotent stem cells (PSCs), including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), hold great potential for research and application in regenerative medicine and drug discovery. To maximize primate PSC potential, a practical system is required for generating desired functional cells and reproducible differentiation techniques. Much progress regarding their culture systems has been reported to date; however, better methods would still be required for their practical use, particularly in industrial and clinical fields. Here we report a new single-cell and feeder-free culture system for primate PSCs, the key feature of which is an originally formulated serum-free medium containing FGF and activin. In this culture system, cynomolgus monkey ESCs can be passaged many times by single-cell dissociation with traditional trypsin treatment and can be propagated with a high proliferation rate as a monolayer without any feeder cells; further, typical PSC properties and genomic stability can be retained. In addition, it has been demonstrated that monkey ESCs maintained in the culture system can be used for various experiments such as in vitro differentiation and gene manipulation. Thus, compared with the conventional culture system, monkey ESCs grown in the aforementioned culture system can serve as a cell source with the following practical advantages: simple, stable, and easy cell maintenance; gene manipulation; cryopreservation; and desired differentiation. We propose that this culture system can serve as a reliable platform to prepare primate PSCs useful for future research and application.
-
Stimulation of the proliferation of hemopoietic stem cells in irradiated bone marrow cell culture
International Nuclear Information System (INIS)
Mori, K.J.; Izumi, H.; Seto, A.
1981-01-01
Long-term hemopoiesis was established in bone marrow cell culture in vitro. This culture was shown to support the recovery proliferation of hemopoietic stem cells completely in vitro after irradiation. Hemopoietic stem cells were stimulated into proliferation in culture when normal bone marrow cells were overlayed on top of the irradiated adherent cell colonies. These results indicate that proliferation and differentiation of hemopoietic stem cells in vitro are also supported by stromahemopoietic cell interactions
-
Taylor, Peter Charles
2011-12-01
John Settlage's article— Counterstories from White Mainstream Preservice Teachers: Resisting the Master Narrative of Deficit by Default—outlines his endeavour to enable pre-service teachers to develop culturally responsive science teaching identities for resisting the master narrative of deficit thinking when confronted by the culturally different `other.' Case study results are presented of the role of counterstories in enabling five pre-service teachers to overcome deficit thinking. In this forum, Philip Moore, a cultural anthropologist and university professor, deepens our understanding of the power and significance of counterstories as an educational tool for enabling students to deconstruct oppressive master narratives. Jill Slay, dean of a science faculty, examines her own master narrative about the compatibility of culturally similar academics and graduate students, and finds it lacking. But first, I introduce this scholarship with background notes on the critical paradigm and its adversary, the grand narrative of science education, following which I give an appreciative understanding of John's pedagogical use of counterstories as a transformative strategy for multi-worldview science teacher education.
-
Cell Culture in Microgravity: Opening the Door to Space Cell Biology
Pellis, Neal R.; Dawson, David L. (Technical Monitor)
1999-01-01
Adaptational response of human cell populations to microgravity is investigated using simulation, short-term Shuttle experiments, and long-term microgravity. Simulation consists of a clinostatically-rotated cell culture system. The system is a horizontally-rotated cylinder completely filled with culture medium. Low speed rotation results in continuous-fall of the cells through the fluid medium. In this setting, cells: 1) aggregate, 2) propagate in three dimensions, 3) synthesize matrix, 4) differentiate, and 5) form sinusoids that facilitate mass transfer. Space cell culture is conducted in flight bioreactors and in static incubators. Cells grown in microgravity are: bovine cartilage, promyelocytic leukemia, kidney proximal tubule cells, adrenal medulla, breast and colon cancer, and endothelium. Cells were cultured in space to test specific hypotheses. Cartilage cells were used to determine structural differences in cartilage grown in space compared to ground-based bioreactors. Results from a 130-day experiment on Mir revealed that cartilage grown in space was substantially more compressible due to insufficient glycosaminoglycan in the matrix. Interestingly, earth-grown cartilage conformed better to the dimensions of the scaffolding material, while the Mir specimens were spherical. The other cell populations are currently being analyzed for cell surface properties, gene expression, and differentiation. Results suggest that some cells spontaneously differentiate in microgravity. Additionally, vast changes in gene expression may occur in response to microgravity. In conclusion, the transition to microgravity may constitute a physical perturbation in cells resulting in unique gene expressions, the consequences of which may be useful in tissue engineering, disease modeling, and space cell biology.
-
Trischler, Helmuth; Carson, Cathryn; Kojevnikov, Alexei
2008-12-01
'"Forman thesis', published in 1971, argued for a historical linkage among the intellectual atmosphere of Weimar Germany, popular revolts against determinism and materialism, and the creation of the revolutionary new theory of quantum mechanics. Paul Forman's long essay on "Weimar Culture" has shaped research agendas in numerous fields, from the history and philosophy of physics to German history to the sociology of scientific knowledge. Despite its status as a classic and its transformative effect, Weimar Culture has always inspired as much critique as assent. In particular in the history of science, cohorts of students and two generations of scholars have debated the Forman thesis as a conceptual tool for linking scientific change with cultural processes. The Forman thesis raises critical questions for both the ongoing debates over cultural approaches to the history of science and the burgeoning newer scholarship on physics in and beyond Weimar Germany. Exploring these implications has been the aim of a transnational project of the three authors of this article which sheds some light on these debates and briefly introduces the following papers of this special issue devoted to Paul Forman and his seminal works in the history of science.
-
How do culture media influence in vitro perivascular cell behavior?
Huber, Birgit; Volz, Ann-Cathrin; Kluger, Petra Juliane
2015-12-01
Perivascular cells are multilineage cells located around the vessel wall and important for wall stabilization. In this study, we evaluated a stem cell media and a perivascular cell-specific media for the culture of primary perivascular cells regarding their cell morphology, doubling time, stem cell properties, and expression of cell type-specific markers. When the two cell culture media were compared to each other, perivascular cells cultured in the stem cell medium had a more elongated morphology and a faster doubling rate and cells cultured in the pericyte medium had a more typical morphology, with several filopodia, and a slower doubling rate. To evaluate stem cell properties, perivascular cells, CD146(-) cells, and mesenchymal stem cells (MSCs) were differentiated into the adipogenic, osteogenic, and chondrogenic lineages. It was seen that perivascular cells, as well as CD146(-) cells and MSCs, cultured in stem cell medium showed greater differentiation than cells cultured in pericyte-specific medium. The expression of pericyte-specific markers CD146, neural/glial antigen 2 (NG2), platelet-derived growth factor receptor-β (PDGFR-β), myosin, and α-smooth muscle actin (α-SMA) could be found in both pericyte cultures, as well as to varying amounts in CD146(-) cells, MSCs, and endothelial cells. The here presented work shows that perivascular cells can adapt to their in vitro environment and cell culture conditions influence cell functionality, such as doubling rate or differentiation behavior. Pericyte-specific markers were shown to be expressed also from cells other than perivascular cells. We can further conclude that CD146(+) perivascular cells are inhomogeneous cell population probably containing stem cell subpopulations, which are located perivascular around capillaries. © 2015 International Federation for Cell Biology.
-
Biogelx: Cell Culture on Self-Assembling Peptide Gels.
Harper, Mhairi M; Connolly, Michael L; Goldie, Laura; Irvine, Eleanore J; Shaw, Joshua E; Jayawarna, Vineetha; Richardson, Stephen M; Dalby, Matthew J; Lightbody, David; Ulijn, Rein V
2018-01-01
Aromatic peptide amphiphiles can form self-supporting nanostructured hydrogels with tunable mechanical properties and chemical compositions. These hydrogels are increasingly applied in two-dimensional (2D) and three-dimensional (3D) cell culture, where there is a rapidly growing need to store, grow, proliferate, and manipulate naturally derived cells within a hydrated, 3D matrix. Biogelx Limited is a biomaterials company, created to commercialize these bio-inspired hydrogels to cell biologists for a range of cell culture applications. This chapter describes methods of various characterization and cell culture techniques specifically optimized for compatibility with Biogelx products.
-
Gallagher, James J., Ed.; Dawson, George, Ed.
The impact of cultural background on science learning is explored in this compilation of papers and reports from an inter-American Seminar on science education. For the purposes of enriching science program planning, teacher education, research, and practice in the schools, varying ideas are offered on the effects of cultural background on science…
-
Cole, Mikel Walker
2015-12-01
This response to Marie Paz Morales' "Influence of culture and language sensitive physics on science attitude achievement" explores the ideas of culturally responsive pedagogy and critical literacy to examine some implications for culturally responsive science instruction implicit in the original manuscript.
-
Pursuing Darwin's curious parallel: Prospects for a science of cultural evolution.
Mesoudi, Alex
2017-07-24
In the past few decades, scholars from several disciplines have pursued the curious parallel noted by Darwin between the genetic evolution of species and the cultural evolution of beliefs, skills, knowledge, languages, institutions, and other forms of socially transmitted information. Here, I review current progress in the pursuit of an evolutionary science of culture that is grounded in both biological and evolutionary theory, but also treats culture as more than a proximate mechanism that is directly controlled by genes. Both genetic and cultural evolution can be described as systems of inherited variation that change over time in response to processes such as selection, migration, and drift. Appropriate differences between genetic and cultural change are taken seriously, such as the possibility in the latter of nonrandomly guided variation or transformation, blending inheritance, and one-to-many transmission. The foundation of cultural evolution was laid in the late 20th century with population-genetic style models of cultural microevolution, and the use of phylogenetic methods to reconstruct cultural macroevolution. Since then, there have been major efforts to understand the sociocognitive mechanisms underlying cumulative cultural evolution, the consequences of demography on cultural evolution, the empirical validity of assumed social learning biases, the relative role of transformative and selective processes, and the use of quantitative phylogenetic and multilevel selection models to understand past and present dynamics of society-level change. I conclude by highlighting the interdisciplinary challenges of studying cultural evolution, including its relation to the traditional social sciences and humanities.
-
Engaging Karen refugee students in science learning through a cross-cultural learning community
Harper, Susan G.
2017-02-01
This research explored how Karen (first-generation refugees from Burma) elementary students engaged with the Next Generation Science Standards (NGSS) practice of constructing scientific explanations based on evidence within the context of a cross-cultural learning community. In this action research, the researcher and a Karen parent served as co-teachers for fourth- and fifth-grade Karen and non-Karen students in a science and culture after-school programme in a public elementary school in the rural southeastern United States. Photovoice provided a critical platform for students to create their own cultural discourses for the learning community. The theoretical framework of critical pedagogy of place provided a way for the learning community to decolonise and re-inhabit the learning spaces with knowledge they co-constructed. Narrative analysis of video transcripts of the after-school programme, ethnographic interviews, and focus group discussions from Photovoice revealed a pattern of emerging agency by Karen students in the scientific practice of constructing scientific explanations based on evidence and in Karen language lessons. This evidence suggests that science learning embedded within a cross-cultural learning community can empower refugee students to construct their own hybrid cultural knowledge and leverage that knowledge to engage in a meaningful way with the epistemology of science.
-
Garvin, Brittany A.
There have been numerous calls and efforts made to provide states, school districts, and communities needed financial support to increase and enhance access to and opportunities in Science, Technology, Engineering, and Math (STEM) related disciplines for marginalized populations (Tyson, Lee, & Hanson, 2007; Caldwell & Siwatu, 2003). As the challenge to better educate students of color and poor students intensifies, the need to provide equitable science learning experiences for all students aimed at scientific literacy and STEM also becomes critical. Thus the need to provide summer science enrichment programs where students engage in scientific experimentation, investigation, and critical thinking are vital to helping students who have been traditionally marginalized achieve success in school science and enter the science career pipeline. This mixed methods study examined the impact of a culturally responsive approach on student attitudes, interests in science education and STEM careers, and basic science content knowledge before and after participation in an upward bound summer program. Quantitative results indicated using a culturally responsive approach to teach science in an informal learning space significantly increases student achievement. Students receiving culturally responsive science instruction exhibited statistically significant increases in their posttest science scores compared to pretest science scores, M = 0.376, 95% CI [0.266, 0.487], t (10) = 7.610, p < 0.001. Likewise, students receiving culturally responsive science instruction had a significantly higher interest in science (M = 1.740, SD = 0.548) and STEM careers, M = 0.597, 95% CI [0.276, 0.919], p = 0.001. The qualitative data obtained in this study sought to gain a more in-depth understanding of the impact of a culturally responsive approach on students' attitudes, interests in science and STEM careers. Findings suggest providing students the opportunity to do and learn science utilizing a
-
Cardiac Cells Beating in Culture: A Laboratory Exercise
Weaver, Debora
2007-01-01
This article describes how to establish a primary tissue culture, where cells are taken directly from an organ of a living animal. Cardiac cells are taken from chick embryos and transferred to culture dishes. These cells are not transformed and therefore have a limited life span. However, the unique characteristics of cardiac cells are maintained…
-
Wang, Ya-Ling; Liang, Jyh-Chong; Tsai, Chin-Chung
2018-04-01
Science learning self-efficacy could be regarded as a multi-factor belief which comprises different aspects such as cognitive skills, practical work, and everyday application. However, few studies have investigated the relationships among these factors that compose science learning self-efficacy. Also, culture may play an important role in explaining the relationships among these factors. Accordingly, this study aimed to investigate cultural differences in science learning self-efficacy and examine the relationships within factors constituting science learning self-efficacy by adopting a survey instrument for administration to students in the U.S. and Taiwan. A total of 218 university students (62.40% females) were surveyed in the U.S.A, and 224 university students (49.10% females) in Taiwan were also invited to take part in the study. The results of the structural equation modelling revealed cultural differences in the relationships among the factors of science learning self-efficacy. It was found that U.S. students' confidence in their ability to employ higher-order cognitive skills tended to promote their confidence in their ability to accomplish practical work, strengthening their academic self-efficacy. However, the aforementioned mediation was not found for the Taiwanese participants.
-
Improved endothelial cell seeding with cultured cells and fibronectin-coated grafts
International Nuclear Information System (INIS)
Seeger, J.M.; Klingman, N.
1985-01-01
A possible approach to the low seeding efficiency of endothelial cells into prosthetic grafts is to increase the number of cells to be seeded in cell culture and improve seeding efficiency by graft precoating with fibronectin. The effect of cell culture on cell adhesion is unknown, however, and fibronectin also binds fibrin, which may increase the thrombogenicity of the graft luminal surface. To investigate these questions, freshly harvested canine jugular vein endothelial cells from six animals and similar cells harvested from six primary and eight secondary cell cultures were labeled with 111 Indium and seeded into 5 cm, 4 mm PTFE grafts coated with fibronectin, using similar uncoated PTFE grafts as controls. Platelet accumulation and distribution on six similar coated and uncoated grafts placed in canine carotid, external jugular arterial venous shunts for 2 hr were also determined using autogenous 111 Indium-labeled platelets. Significant differences between group means were determined using the paired Student's t test. Results reveal that seeding efficiency is significantly better in all groups of coated grafts compared to uncoated grafts (P less than 0.01). Cells derived from cell culture also had significantly higher seeding efficiencies than freshly harvested cells when seeded into coated grafts (P less than 0.05) and tended to have higher seeding efficiencies than harvested cells when seeded into uncoated grafts (P = 0.53). Fibronectin coating increased mean platelet accumulation on the entire graft luminal surface, but not to a statistically significant degree (P greater than 0.1). Whether this increased seeding efficiency will improve graft endothelialization remains to be investigated
-
Dynamic cell culture system: a new cell cultivation instrument for biological experiments in space
Gmunder, F. K.; Nordau, C. G.; Tschopp, A.; Huber, B.; Cogoli, A.
1988-01-01
The prototype of a miniaturized cell cultivation instrument for animal cell culture experiments aboard Spacelab is presented (Dynamic cell culture system: DCCS). The cell chamber is completely filled and has a working volume of 200 microliters. Medium exchange is achieved with a self-powered osmotic pump (flowrate 1 microliter h-1). The reservoir volume of culture medium is 230 microliters. The system is neither mechanically stirred nor equipped with sensors. Hamster kidney (Hak) cells growing on Cytodex 3 microcarriers were used to test the biological performance of the DCCS. Growth characteristics in the DCCS, as judged by maximal cell density, glucose consumption, lactic acid secretion and pH, were similar to those in cell culture tubes.
-
Quantitative volumetric Raman imaging of three dimensional cell cultures
Kallepitis, Charalambos; Bergholt, Mads S.; Mazo, Manuel M.; Leonardo, Vincent; Skaalure, Stacey C.; Maynard, Stephanie A.; Stevens, Molly M.
2017-01-01
in conventional cell culture systems and mesenchymal stem cells inside biomimetic hydrogels that supplied a 3D cell culture environment. We demonstrate visualization and quantification of fine details in cell shape, cytoplasm, nucleus, lipid bodies
-
System-level modeling and simulation of the cell culture microfluidic biochip ProCell
DEFF Research Database (Denmark)
Minhass, Wajid Hassan; Pop, Paul; Madsen, Jan
2010-01-01
Microfluidic biochips offer a promising alternative to a conventional biochemical laboratory. There are two technologies for the microfluidic biochips: droplet-based and flow-based. In this paper we are interested in flow-based microfluidic biochips, where the liquid flows continuously through pre......-defined micro-channels using valves and pumps. We present an approach to the system-level modeling and simulation of a cell culture microfluidic biochip called ProCell, Programmable Cell Culture Chip. ProCell contains a cell culture chamber, which is envisioned to run 256 simultaneous experiments (viewed...
-
DEFF Research Database (Denmark)
Al Atraktchi, Fatima Al-Zahraa; Bakmand, Tanya; Rømer Sørensen, Ane
The brain is the center of the nervous system, where serious neurodegenerative diseases such as Parkinson’s, Alzheimer’s and Huntington’s are products of functional loss in the neural cells (1). Typical techniques used to investigate these diseases lack precise control of the cellular surroundings......, in addition to isolating the neural tissue from nutrient delivery and to creating unwanted gradients (2). This means that typical techniques used to investigate neurodegenerative diseases cannot mimic in vivo conditions, as closely as desired. We have developed a novel microfluidic system for culturing PC12...... cells, neuronal cells, astrocytes cultures and brain slices. The microfluidic system provides efficient nutrient delivery, waste removal, access to oxygen, fine control over the neurochemical environment and access to modern microscopy. Additionally, the setup consists of an in vitro culturing...
-
A Survey of Cultural Infrastructure and Performance in Medical Sciences Universities of Iran
Directory of Open Access Journals (Sweden)
Mahmood Feizi
2015-08-01
Full Text Available Background and objectives: Recently, the role of universities in developing and education of culture is considered increasingly but Iranian universities have great distance in achieving the desired objectives in this context. So, this study aimed to survey the cultural infrastructure and performance in medical sciences universities of Iran. Material and Methods: This is a cross-sectional study that was done using researcher-made checklist which its face and content validity were approved by the cultural experts' opinion via statistical indicators. The study was conducted in census method by responses of 25 managers of cultural affairs in medical sciences universities of Iran. The obtained data were analyzed descriptively and results were reported as frequency (percentages for qualitative and mean (standard deviation for quantitative variable. Results: The study results were presented in four areas: “the general status of universities in cultural affairs”, “cultural facilities of the universities”, “the activity of cultural organizations and publications in universities” and “performance of cultural deputies”. The results showed that although there are considerable strengths, the significant weaknesses are evident in all areas. The results of the present study were focused solely on the quantity of functions, and quality evaluation of each activity requires special attention and further investigations and interventions. Conclusion: Researchers hope that the authorities and planners use the results of this study and similar studies especially in quality of cultural practices of universities and move towards improving the status of culture in medical sciences universities in developing Iranian-Islamic culture.
-
Maintenance of mesenchymal stem cells culture due to the cells with reduced attachment rate
Directory of Open Access Journals (Sweden)
Shuvalova N. S.
2013-01-01
Full Text Available Aim. The classic detachment techniques lead to changes in cells properties. We offer a simple method of cultivating the population of cells that avoided an influence on the surface structures. Methods. Mesenchymal stem cells (MSC from human umbilical cord matrix were obtained and cultivated in standard conditions. While substituting the culture media by a fresh portion, the conditioned culture medium, where the cells were maintained for three days, was transferred to other culture flacks with addition of serum and growth factors. Results. In the flacks, one day after medium transfer, we observed attached cells with typical MSC morphology. The cultures originated from these cells had the same rate of surface markers expression and clonogenic potential as those replated by standard methods. Conclusions. MSC culture, derived by preserving the cells with reduced attachment ability, actually has the properties of «parent» passage. Using this method with accepted techniques of cells reseeding would allow maintaining the cells that avoided an impact on the cell surface proteins.
-
Lee, Sung-Il; Ko, Youngkyung; Park, Jun-Beom
2017-09-01
Three-dimensional cell culture systems provide a convenient in vitro model for the study of complex cell-cell and cell-matrix interactions in the absence of exogenous substrates. The current study aimed to evaluate the osteogenic differentiation potential of gingiva-derived stem cells cultured in two-dimensional or three-dimensional systems. To the best of our knowledge, the present study is the first to compare the growth of gingiva-derived stem cells in monolayer culture to a three-dimensional culture system with microwells. For three-dimensional culture, gingiva-derived stem cells were isolated and seeded into polydimethylsiloxane-based concave micromolds. Alkaline phosphatase activity and alizarin red S staining assays were then performed to evaluate osteogenesis and the degree of mineralization, respectively. Stem cell spheroids had a significantly increased level of alkaline phosphatase activity and mineralization compared with cells from the two-dimensional culture. In addition, an increase in mineralized deposits was observed with an increase in the loading cell number. The results of present study indicate that gingiva-derived stem cell spheroids exhibit an increased osteogenic potential compared with stem cells from two-dimensional culture. This highlights the potential of three-dimensional culture systems using gingiva-derived stem cells for regenerative medicine applications requiring stem cells with osteogenic potential.
-
Cole, Mikel Walker
2015-01-01
This response to Marie Paz Morales' "Influence of culture and language sensitive physics on science attitude achievement" explores the ideas of culturally responsive pedagogy and critical literacy to examine some implications for culturally responsive science instruction implicit in the original manuscript. [For "Influence of…
-
Koban, Robert; Neumann, Markus; Daugs, Aila; Bloch, Oliver; Nitsche, Andreas; Langhammer, Stefan; Ellerbrok, Heinz
2018-02-01
Gefitinib is a specific inhibitor of the epidermal growth factor receptor (EGFR) and FDA approved for treatment of non-small cell lung cancer. In a previous study we could show the in vitro efficacy of gefitinib for treatment of poxvirus infections in monolayer (2D) cultivated cell lines. Permanent cell lines and 2D cultures, however, are known to be rather unphysiological; therefore it is difficult to predict whether determined effective concentrations or the drug efficacy per se are transferable to the in vivo situation. 3D cell cultures, which meanwhile are widely distributed across all fields of research, are a promising tool for more predictive in vitro investigations of antiviral compounds. In this study the spreading of cowpox virus and the antiviral efficacy of gefitinib were analyzed in primary human keratinocytes (NHEK) grown in a novel 3D extracellular matrix-based cell culture model and compared to the respective monolayer culture. 3D-cultivated NHEK grew in a polarized and thus a more physiological manner with altered morphology and close cell-cell contact. Infected cultures showed a strongly elevated sensitivity towards gefitinib. EGFR phosphorylation, cell proliferation, and virus replication were significantly reduced in 3D cultures at gefitinib concentrations which were at least 100-fold lower than those in monolayer cultures and well below the level of cytotoxicity. Our newly established 3D cell culture model with primary human cells is an easy-to-handle alternative to conventional monolayer cell cultures and previously described more complex 3D cell culture systems. It can easily be adapted to other cell types and a broad spectrum of viruses for antiviral drug screening and many other aspects of virus research under more in vivo-like conditions. In consequence, it may contribute to a more targeted realization of necessary in vivo experiments. Copyright © 2017 Elsevier B.V. All rights reserved.
-
SCIENCE FICTION IN HISTORICAL AND CULTURAL LITERARY DISCOURSE
Directory of Open Access Journals (Sweden)
Simona Siderevičiūtė
2014-04-01
Full Text Available This work intends to complement literary studies in science fiction. It discusses the history of global science fiction, overviews the most characteristic features of its historical periods, and provides an introduction to Lithuanian science fiction, indicating its main features and topics. In the context of culture, science fiction is often defined as a literary genre with the emphasis on its nature as fiction. Only rarely are the history of the origin of science fiction, its variations, and the pioneers of science fiction whose works are still highly valued taken into account. Science fiction is often criticized through the filter of preconceived ideas that consider this type of literature to be “frivolous.” This article discusses the possible reasons for such an approach. In Lithuania, this genre is still associated only with pop literature, and its expression cannot yet equal the works of foreign authors. The basic classical motifs of global science fiction found in Lithuanian science fiction include: representatives of extraterrestrial civilizations and human contact with them, scientists and inventors, agents of military institutions, and space travel. Lithuanian science fiction writers follow the traditions of global science fiction when using these classical motifs; however, a general lack of original and individual themes, motifs, and manifestations may be observed.
-
Relation of Astronomy to other Sciences, Culture and Society
Harutyunian, H. A.; Mickaelian, A. M.; Farmanyan, S. V.
2015-07-01
The book contains the Proceedings of XIII Annual Meeting of the Armenian Astronomical Society "Relation of Astronomy to other Sciences, Culture and Society". It consists of 9 main sections: "Introductory", "Astronomy and Philosophy", "Astrobiology", "Space-Earth Connections", "Astrostatistics and Astroinformatics", "Astronomy and Culture, Astrolinguistics", "Archaeoastronomy", "Scientific Tourism and Scientific Journalism", and "Armenian Astronomy". The book may be interesting to astronomers, philosophers, biologists, culturologists, linguists, historians, archaeologists and to other specialists, as well as to students.
-
Neuroanthropology: a humanistic science for the study of the culture-brain nexus.
Domínguez Duque, Juan F; Turner, Robert; Lewis, E Douglas; Egan, Gary
2010-06-01
In this article, we argue that a combined anthropology/neuroscience field of enquiry can make a significant and distinctive contribution to the study of the relationship between culture and the brain. This field, which can appropriately be termed as neuroanthropology, is conceived of as being complementary to and mutually informative with social and cultural neuroscience. We start by providing an introduction to the culture concept in anthropology. We then present a detailed characterization of neuroanthropology and its methods and how they relate to the anthropological understanding of culture. The field is described as a humanistic science, that is, a field of enquiry founded on the perceived epistemological and methodological interdependence of science and the humanities. We also provide examples that illustrate the proposed methodological model for neuroanthropology. We conclude with a discussion about specific contributions the field can make to the study of the culture-brain nexus.
-
Mesenchymal stem cells enhance the metastasis of 3D-cultured hepatocellular carcinoma cells
International Nuclear Information System (INIS)
Liu, Chang; Liu, Yang; Xu, Xiao-xi; Guo, Xin; Sun, Guang-wei; Ma, Xiao-jun
2016-01-01
Accumulating evidences have demonstrated that mesenchymal stem cells (MSC) could be recruited to the tumor microenvironment. Umbilical cord mesenchymal stem cells (UCMSC) were attractive vehicles for delivering therapeutic agents against cancer. Nevertheless, the safety of UCMSC in the treatment of tumors including hepatocellular carcinoma (HCC) was still undetermined. In this study, an in vitro co-culture system was established to evaluate the effect of UCMSC on the cell growth, cancer stem cell (CSC) characteristics, drug resistance, metastasis of 3D-cultured HCC cells, and the underlying mechanism was also investigated. It was found that after co-cultured with UCMSC, the metastatic ability of 3D-cultured HCC cells was significantly enhanced as indicated by up-regulation of matrix metalloproteinase (MMP), epithelial-mesenchymal transition (EMT)-related genes, and migration ability. However, cell growth, drug resistance and CSC-related gene expression of HCC cells were not affected by UCMSC. Moreover, EMT was reversed, MMP-2 expression was down-regulated, and migration ability of HCC cell was significantly inhibited when TGF-β receptor inhibitor SB431542 was added into the co-culture system. Therefore, these data indicated that UCMSC could significantly enhance the tumor cell metastasis, which was due to the EMT of HCC cells induced by TGF-β. The online version of this article (doi:10.1186/s12885-016-2595-4) contains supplementary material, which is available to authorized users
-
Bonazza, Camila; Andrade, Sheila Siqueira; Sumikawa, Joana Tomomi; Batista, Fabrício Pereira; Paredes-Gamero, Edgar J; Girão, Manoel J B C; Oliva, Maria Luiza V; Castro, Rodrigo Aquino
2016-01-01
Cell culture is considered the standard media used in research to emulate the in vivo cell environment. Crucial in vivo experiments cannot be conducted in humans and depend on in vitro methodologies such as cell culture systems. However, some procedures involving the quality control of cells in culture have been gradually neglected by failing to acknowledge that primary cells and cell lines change over time in culture. Thus, we report methods based on our experience for monitoring primary cell culture of human myometrial cells derived from uterine leiomyoma. We standardized the best procedure of tissue dissociation required for the study of multiple genetic marker systems that include species-specific antigens, expression of myofibroblast or myoblast markers, growth curve, serum deprivation, starvation by cell cycle synchronization, culture on collagen coated plates, and 17 β-estradiol (E2) and progesterone (P4) effects. The results showed that primary myometrial cells from patients with uterine leiomyoma displayed myoblast phenotypes before and after in vitro cultivation, and leiomyoma cells differentiated into mature myocyte cells under the appropriate differentiation-inducing conditions (serum deprivation). These cells grew well on collagen coated plates and responded to E2 and P4, which may drive myometrial and leiomyoma cells to proliferate and adhere into a focal adhesion complex involvement in a paracrine manner. The establishment of these techniques as routine procedures will improve the understanding of the myometrial physiology and pathogenesis of myometrium-derived diseases such as leiomyoma. Mimicking the in vivo environment of fibrotic conditions can prevent false results and enhance results that are based on cell culture integrity.
-
The Importance of Cultural Heritage in Earth Science
Avvisati, Gala; Di Vito, Mauro; Marotta, Enrica; Sangianantoni, Agata; Peluso, Rosario; de Vita, Sandro; Nave, Rosella; Vertechi, Enrico; De Natale, Giuseppe; Ghilardi, Massimo
2016-04-01
In recent years the Earth Sciences community is facing the need to achieve a more effective and efficient dissemination of its scientific culture. There is now a growing needing to integrate the use of "traditional" dissemination media of cultural heritage with the new digital technologies. Getting people involved in geoheritage site's activities represents a crucial issue in order to better communicate and increase the collective awareness of natural hazards, risk, and environmental change. The Reale Osservatorio Vesuviano (ROV) which is part of the Istituto Nazionale di Geofisica e Vulcanologia (INGV), owns collections unique in their combination of scientific, historical and artistic importance. The long history of ROV is extensively documented in its collections. This heritage - of great scientific and cultural value and unique for its abundance and variety - tells the story of the first observatory in the world, closely linked to the activity of Vesuvius, and the commitment of many scientists who dedicated their lives to study the volcano. The collections include: a) old books on volcanological matters, b) collection of rocks, minerals, volcanic ash and other materials from historical eruptions of Vesuvius, c) recordings on smoked paper of Vesuvius seismic activity from 1915 until 1970, d) scientific instruments, e) geological and geomorphological maps and models, f) vintage photographs and filmed sequences of eruptions, g) gouaches of Vesuvius and h) lava medals. The exposition of these collections, improved with the new digital contents, may trace new and unexplored routes for the dissemination of Earth Sciences related culture. The ethical duty of the ROV is the creation of an universal identity by taking a picture of the evolution of the society through the training of the culture of seismic and volcanic risk. A disappearance of its heritage could represent an huge impoverishment of its community: the ROV carries in fact the cultural identity of the
-
Kasperowski, Dick; Hillman, Thomas
2018-05-01
In the past decade, some areas of science have begun turning to masses of online volunteers through open calls for generating and classifying very large sets of data. The purpose of this study is to investigate the epistemic culture of a large-scale online citizen science project, the Galaxy Zoo, that turns to volunteers for the classification of images of galaxies. For this task, we chose to apply the concepts of programs and antiprograms to examine the 'essential tensions' that arise in relation to the mobilizing values of a citizen science project and the epistemic subjects and cultures that are enacted by its volunteers. Our premise is that these tensions reveal central features of the epistemic subjects and distributed cognition of epistemic cultures in these large-scale citizen science projects.
-
Radiosensitivity of normal human epidermal cells in culture
International Nuclear Information System (INIS)
Dover, R.; Potten, C.S.
1983-01-01
Using an in vitro culture system the authors have derived #betta#-radiation survival curves over a dose range 0-8 Gy for the clonogenic cells of normal human epidermis. The culture system used allows the epidermal cells to stratify and form a multi-layered sheet of keratinizing cells. The cultures appear to be a very good model for epidermis in vivo. The survival curves show a population which is apparently more sensitive than murine epidermis in vivo. It remains unclear whether this is an intrinsic difference between the species or is a consequence of the in vitro cultivation of the human cells. (author)
-
Yamashita, Tomoko; Miyamoto, Yuki; Bando, Yoshio; Ono, Takashi; Kobayashi, Sakurako; Doi, Ayano; Araki, Toshihiro; Kato, Yosuke; Shirakawa, Takayuki; Suzuki, Yutaka; Yamauchi, Junji; Yoshida, Shigetaka; Sato, Naoya
2017-01-01
Oligodendrocytes myelinate axons and form myelin sheaths in the central nervous system. The development of therapies for demyelinating diseases, including multiple sclerosis and leukodystrophies, is a challenge because the pathogenic mechanisms of disease remain poorly understood. Primate pluripotent stem cell-derived oligodendrocytes are expected to help elucidate the molecular pathogenesis of these diseases. Oligodendrocytes have been successfully differentiated from human pluripotent stem cells. However, it is challenging to prepare large amounts of oligodendrocytes over a short amount of time because of manipulation difficulties under conventional primate pluripotent stem cell culture methods. We developed a proprietary dissociated monolayer and feeder-free culture system to handle pluripotent stem cell cultures. Because the dissociated monolayer and feeder-free culture system improves the quality and growth of primate pluripotent stem cells, these cells could potentially be differentiated into any desired functional cells and consistently cultured in large-scale conditions. In the current study, oligodendrocyte progenitor cells and mature oligodendrocytes were generated within three months from monkey embryonic stem cells. The embryonic stem cell-derived oligodendrocytes exhibited in vitro myelinogenic potency with rat dorsal root ganglion neurons. Additionally, the transplanted oligodendrocyte progenitor cells differentiated into myelin basic protein-positive mature oligodendrocytes in the mouse corpus callosum. This preparative method was used for human induced pluripotent stem cells, which were also successfully differentiated into oligodendrocyte progenitor cells and mature oligodendrocytes that were capable of myelinating rat dorsal root ganglion neurons. Moreover, it was possible to freeze, thaw, and successfully re-culture the differentiating cells. These results showed that embryonic stem cells and human induced pluripotent stem cells maintained in a
-
Gu, Wenyu; Sun, Wei; Guo, Changcheng; Yan, Yang; Liu, Min; Yao, Xudong; Yang, Bin; Zheng, Junhua
2015-07-01
Although emerging evidence demonstrates increased circulating endothelial progenitor cells in patients with solid tumors, to our knowledge it is still unknown whether such cells can be cultured from patients with highly angiogenic renal cell carcinoma. We cultured and characterized circulating endothelial progenitor cells from patients with renal cell carcinoma. The circulating endothelial progenitor cell level (percent of CD45(-)CD34(+) VEGF-R2(+) cells in total peripheral blood mononuclear cells) was quantified in 47 patients with renal cell carcinoma and 40 healthy controls. Peripheral blood mononuclear cells were then isolated from 33 patients with renal cell carcinoma and 30 healthy controls to culture and characterize circulating endothelial progenitor cells. The circulating endothelial progenitor cell level was significantly higher in patients with renal cell carcinoma than in healthy controls (0.276% vs 0.086%, p cells first emerged significantly earlier in patient than in control preparations (6.72 vs 14.67 days, p culture success rate (87.8% vs 40.0% of participants) and the number of colonies (10.06 vs 1.83) were significantly greater for patients than for controls (each p cell level correlated positively with the number of patient colonies (r = 0.762, p Cells cultured from patients and controls showed a similar growth pattern, immunophenotype, ability to uptake Ac-LDL and bind lectin, and form capillary tubes in vitro. However, significantly more VEGF-R2(+) circulating endothelial progenitor cells were found in preparations from patients with renal cell carcinoma than from healthy controls (21.1% vs 13.4%, p cell colonies, a higher cell culture success rate and more colonies were found for patients with renal cell carcinoma than for healthy controls. Results indicate the important significance of VEGF-R2(+) circulating endothelial progenitors in patients with renal cell carcinoma. Copyright © 2015 American Urological Association Education and Research
-
The impact of cell culture equipment on energy loss.
Davies, Lleucu B; Kiernan, Michael N; Bishop, Joanna C; Thornton, Catherine A; Morgan, Gareth
2014-01-01
Light energy of discrete wavelengths supplied via lasers and broadband intense pulsed light have been used therapeutically for many years. In vitro models complement clinical studies, especially for the elucidation of underlying mechanisms of action. Clarification that light energy reaches the cells is necessary when developing protocols for the treatment of cells using in vitro models. Few studies report on energy loss in cell culture equipment. The ability of energy from light with therapeutic potential to reach cells in culture needs to be determined; this includes determining the proportion of light energy lost within standard cell culture media and cell culture vessels. The energy absorption of cell culture media, with/without the pH indicator dye phenol red, and the loss of energy within different plastics and glassware used typically for in vitro cell culture were investigated using intense pulsed light and a yellow pulsed dye laser. Media containing phenol red have a distinctive absorption peak (560 nm) absent in phenol red-free media and restored by the addition of phenol red. For both light sources, energy loss was lowest in standard polystyrene tissue culture flasks or multi-well plates and highest in polypropylene vessels or glass tubes. The effects of phenol red-free media on the absorption of energy varied with the light source used. Phenol red-free media are the media of choice; polystyrene vessels with flat surfaces such as culture flasks or multi-well plates should be used in preference to polypropylene or glass vessels.
-
Surface modified alginate microcapsules for 3D cell culture
Chen, Yi-Wen; Kuo, Chiung Wen; Chueh, Di-Yen; Chen, Peilin
2016-06-01
Culture as three dimensional cell aggregates or spheroids can offer an ideal platform for tissue engineering applications and for pharmaceutical screening. Such 3D culture models, however, may suffer from the problems such as immune response and ineffective and cumbersome culture. This paper describes a simple method for producing microcapsules with alginate cores and a thin shell of poly(L-lysine)-graft-poly(ethylene glycol) (PLL-g-PEG) to encapsulate mouse induced pluripotent stem (miPS) cells, generating a non-fouling surface as an effective immunoisolation barrier. We demonstrated the trapping of the alginate microcapsules in a microwell array for the continuous observation and culture of a large number of encapsulated miPS cells in parallel. miPS cells cultured in the microcapsules survived well and proliferated to form a single cell aggregate. Droplet formation of monodisperse microcapsules with controlled size combined with flow cytometry provided an efficient way to quantitatively analyze the growth of encapsulated cells in a high-throughput manner. The simple and cost-effective coating technique employed to produce the core-shell microcapsules could be used in the emerging field of cell therapy. The microwell array would provide a convenient, user friendly and high-throughput platform for long-term cell culture and monitoring.
-
A Simple Hydrophilic Treatment of SU-8 Surfaces for Cell Culturing and Cell Patterning
DEFF Research Database (Denmark)
Wang, Zhenyu; Stangegaard, Michael; Dufva, Hans Martin
2005-01-01
SU-8, an epoxy-based photoresist, widely used in constitution different mTAS systems, is incompatible with mammalian cell adhesion and culture in its native form. Here, we demonstrate a simple, cheap and robust two-step method to render a SU-8 surface hydrophilic and compatible with cell culture........ The contact angle of SU-8 surface was significantly reduced from 90° to 25° after the surface modification. The treated SU-8 surfaces provided a cell culture environment that was comparable with cell culture flask surface in terms of generation time and morphology....
-
DEFF Research Database (Denmark)
Høgh, Mette; Islin, Henrik; Møller, Charlotte
2006-01-01
The interaction between the cell types was simulated in vitro by growing primary cell cultures of human endometrial epithelial cells and trophoblast cells together (co-culture) and separately (control cultures). Gene expression in the cell cultures was compared using the Differential Display method and confirmed...
-
Establishment of automated culture system for murine induced pluripotent stem cells
Directory of Open Access Journals (Sweden)
Koike Hiroyuki
2012-11-01
Full Text Available Abstract Background Induced pluripotent stem (iPS cells can differentiate into any cell type, which makes them an attractive resource in fields such as regenerative medicine, drug screening, or in vitro toxicology. The most important prerequisite for these industrial applications is stable supply and uniform quality of iPS cells. Variation in quality largely results from differences in handling skills between operators in laboratories. To minimize these differences, establishment of an automated iPS cell culture system is necessary. Results We developed a standardized mouse iPS cell maintenance culture, using an automated cell culture system housed in a CO2 incubator commonly used in many laboratories. The iPS cells propagated in a chamber uniquely designed for automated culture and showed specific colony morphology, as for manual culture. A cell detachment device in the system passaged iPS cells automatically by dispersing colonies to single cells. In addition, iPS cells were passaged without any change in colony morphology or expression of undifferentiated stem cell markers during the 4 weeks of automated culture. Conclusions Our results show that use of this compact, automated cell culture system facilitates stable iPS cell culture without obvious effects on iPS cell pluripotency or colony-forming ability. The feasibility of iPS cell culture automation may greatly facilitate the use of this versatile cell source for a variety of biomedical applications.
-
Graham, Phillip W; Kim, Mimi M; Clinton-Sherrod, A Monique; Yaros, Anna; Richmond, Alan N; Jackson, Melvin; Corbie-Smith, Giselle
2016-03-01
Concepts of culture and diversity are necessary considerations in the scientific application of theory generation and developmental processes of preventive interventions; yet, culture and/or diversity are often overlooked until later stages (e.g., adaptation [T3] and dissemination [T4]) of the translational science process. Here, we present a conceptual framework focused on the seamless incorporation of culture and diversity throughout the various stages of the translational science process (T1-T5). Informed by a community-engaged research approach, this framework guides integration of cultural and diversity considerations at each phase with emphasis on the importance and value of "citizen scientists" being research partners to promote ecological validity. The integrated partnership covers the first phase of intervention development through final phases that ultimately facilitate more global, universal translation of changes in attitudes, norms, and systems. Our comprehensive model for incorporating culture and diversity into translational research provides a basis for further discussion and translational science development.
-
Directory of Open Access Journals (Sweden)
mohammad saeed zokaei
2009-11-01
Full Text Available Iranian youth’s leisure culture has been immediately affected by the digital media culture. As a communicative media, cell phone has crossed borders of youth norms and identity; and in addition to facilitating their communication, has changed its patterns. Applying Bourdieu’s concepts of habitus and field, and relied on the qualitative and quantitative data gathered from the mobile youth users, the present study argues that mobile has produced a new field in which youth’s opportunities for leisure, entertainment, communication, and independence have extended. In addition, cell phone has facilitated and compensated for some defects in public sphere, and therefore empowered youth agency, individuality, and power. Despite this strengthening, cell phone does not cross borders of gender and class differences, or the levels of social capital.
-
Directory of Open Access Journals (Sweden)
Camila Bonazza
Full Text Available Cell culture is considered the standard media used in research to emulate the in vivo cell environment. Crucial in vivo experiments cannot be conducted in humans and depend on in vitro methodologies such as cell culture systems. However, some procedures involving the quality control of cells in culture have been gradually neglected by failing to acknowledge that primary cells and cell lines change over time in culture. Thus, we report methods based on our experience for monitoring primary cell culture of human myometrial cells derived from uterine leiomyoma. We standardized the best procedure of tissue dissociation required for the study of multiple genetic marker systems that include species-specific antigens, expression of myofibroblast or myoblast markers, growth curve, serum deprivation, starvation by cell cycle synchronization, culture on collagen coated plates, and 17 β-estradiol (E2 and progesterone (P4 effects. The results showed that primary myometrial cells from patients with uterine leiomyoma displayed myoblast phenotypes before and after in vitro cultivation, and leiomyoma cells differentiated into mature myocyte cells under the appropriate differentiation-inducing conditions (serum deprivation. These cells grew well on collagen coated plates and responded to E2 and P4, which may drive myometrial and leiomyoma cells to proliferate and adhere into a focal adhesion complex involvement in a paracrine manner. The establishment of these techniques as routine procedures will improve the understanding of the myometrial physiology and pathogenesis of myometrium-derived diseases such as leiomyoma. Mimicking the in vivo environment of fibrotic conditions can prevent false results and enhance results that are based on cell culture integrity.
-
Chiu, Mei-Shiu
2010-01-01
The aim of the present study is twofold: (1) to investigate gender differences in the effects of science interest and environmental responsibility on science aspiration and achievement and (2) to explore the relations between cultural supports (macroeconomic and gender equality) and both boys' and girls' tendencies to integrate the aforementioned…
-
How Do Scientists Cross Cultural Borders Between Religion and Science: A Case Study
Barner, Chester A., III
The cultures of science and religion have had different levels of conflict throughout the past several hundred years due in part to the development of the theory of evolution. Although many ideas abound in science education as to the alleviation of this struggle, few studies have examined how scientists who profess religious beliefs deal with this conflict. In general, the study sought to understand the cognitive dynamic of the cultural interaction between the scientific and religious culture within a few individuals. Specifically, the study allowed scientists to explain how they found a measure of compatibility between their faith and their scientific endeavors. Within the boundaries of both the general and specific purposes for the study, the following research question was used: How do college science professors describe the interaction between their faith and their scientific knowledge in reference to their transitioning between a naturalistic or scientific understanding and a super-naturalistic or religious understanding? Three theoretical lenses were used as backdrop to view the cultural interaction. World View (Kearney, 1984), Collateral Learning Theory (Jegede, 1995), and Faith Perspective in relation to the Stages of Faith Theory (Fowler, 1981) constituted the theoretical framework. Because of the qualitative nature of the research, the author used a modified naturalistic paradigm that stressed an emergent quality, grounded categorical design, and a modified case study written format that aided in the understanding of data generated through multiple qualitative methods. Three overlapping themes emerged within the data that offer new insights not only into the complex nature of the conflict but also into the ways scientists themselves find a reason to have faith as well as scientific knowledge. Boundaries based upon a philosophical and world view difference, conflict due to culturally integrative ideas, and cultural bridges without distortion made up the
-
Fleer, Marilyn; Adams, Megan; Gunstone, Richard; Hao, Yijun
2016-01-01
It has been reported that in cross-cultural contexts, Western science content is often not used in everyday practice, and the learning of science is often viewed as difficult and having no social meaning (e.g., Aikenhead & Michell, 2011). It is suggested that the cultural relevance of everyday family practices and Western constructions of…
-
Cross-Cultural Comparisons of Undergraduate Student Views of the Nature of Science
Arino de la Rubia, Leigh S.; Lin, Tzung-Jin; Tsai, Chin-Chung
2014-07-01
Past studies investigating university level students' views of nature of science (NOS) were relatively few and most of them were conducted in Western countries. This paper focuses upon comparing the quantitative patterns in Western (US Caucasian and African-American) and non-Western (Taiwanese) students' views of NOS (VNOS) by adopting a survey instrument. This analysis combined with qualitative data begin to uncover details of potential cultural differences in patterns specifically in the US educational context by comparing Caucasian and African-American student responses to a question from a commonly used assessment of VNOS. Results show different patterns of views along the four dimensions of NOS (social negotiation, invented/creative NOS, cultural impacts, and changing/tentative feature of science) according to student major, student gender, and student ethnicity. These differences and similarities have the potential to impact undergraduate education and underrepresentation of cultural minorities in science careers and call for further research into NOS views in the context of diverse student groups.
-
Salter, Brian; Salter, Charlotte
2013-12-01
Scientific progress in the life sciences is dependent on the governance of tensions between the economic potential of the innovation and the cultural response from society. Ownership of the scientific innovation through patenting is a necessary part of the realization of its economic value yet, in the case of human embryonic stem cell (hESC) science, ownership of the human body and human life may offend fundamental cultural values. In the case of transnational patenting governance by the European Patent Office (EPO) and the European Union (EU), cross-national cultural conflict in the field of hESC science has produced a political demand for a form of governance that can incorporate ethical as well as economic judgements in its decision making. This paper explores how bioethics has responded to this opportunity to establish itself as a form of expert authority for the negotiation and resolution of the cultural conflict. In so doing, it shows how the political struggle that has accompanied this bid for new governance territory has been influenced both by the political tensions between the EPO and EU systems of patenting governance and the resistance of competing experts in law and science to a bioethical presence. Copyright © 2012 Elsevier Ltd. All rights reserved.
-
The Intersection of Identity, Culture and Science Engagement
Strong, LaToya
2016-01-01
Ivã Gurgel, Mauricio Pietrocola, and Graciella Watanabe expand upon the existing literature, which links identity and science engagement. Specifically, the authors focus on ways in which the cultural identities of students relate to their engagement in physics. In doing so, Gurgel, Pietrocola, and Watanabe further build upon the idea that one's…
-
Cell culture density affects the proliferation activity of human adipose tissue stem cells.
Kim, Dae Seong; Lee, Myoung Woo; Ko, Young Jong; Chun, Yong Hoon; Kim, Hyung Joon; Sung, Ki Woong; Koo, Hong Hoe; Yoo, Keon Hee
2016-01-01
In this study, we investigated the effect of cell density on the proliferation activity of human mesenchymal stem cells (MSCs) derived from adipose tissue (AT-MSCs) over time in culture. Passage #4 (P4) and #12 (P12) AT-MSCs from two donors were plated at a density of 200 (culture condition 1, CC1) or 5000 (culture condition 2, CC2) cells cm(-2) . After 7 days of incubation, P4 and P12 AT-MSCs cultured in CC1 were thin and spindle-shaped, whereas those cultured in CC2 had extensive cell-to-cell contacts and an expanded cell volume. In addition, P4 and P12 AT-MSCs in CC1 divided more than three times, while those in CC2 divided less than once on average. Flow cytometric analysis using 5(6)-carboxyfluorescein diacetate N-succinimidyl ester dye showed that the fluorescence intensity of AT-MSCs was lower in CC1 than in CC2. Furthermore, expression of proliferation-associated genes, such as CDC45L, CDC20A and KIF20A, in P4 AT-MSCs was higher in CC1 than in CC2, and this difference was also observed in P12 AT-MSCs. These data demonstrated that cell culture density affects the proliferation activity of MSCs, suggesting that it is feasible to design a strategy to prepare suitable MSCs using specific culture conditions. Copyright © 2016 John Wiley & Sons, Ltd.
-
Nanotechnology, Cell Culture and Tissue Engineering
Directory of Open Access Journals (Sweden)
Kazutoshi Haraguchi
2011-01-01
Full Text Available We have fabricated new types of polymer hydrogels and polymer nanocomposites, i.e., nanocomposite gels (NC gels and soft, polymer nanocomposites (M-NCs: solid, with novel organic/inorganic network structures. Both NC gels and M-NCs were synthesized by in-situ free-radical polymerization in the presence of exfoliated clay platelets in aqueous systems and were obtained in various forms such as film, sheet, tube, coating, etc. and sizes with a wide range of clay contents. Here, disk-like inorganic clay nanoparticles act as multi-functional crosslinkers to form new types of network systems. Both NC gels and M-NCs have extraordinary optical and mechanical properties including ultra-high reversible extensibility, as well as a number of new characteristics relating to optical anisotropy, polymer/clay morphology, biocompatibility, stimuli-sensitive surfaces, micro-patterning, etc. For examples, the biological testing of medical devices, comprised of a sensitization test, an irritation test, an intracutaneous test and an in vitro cytotoxicity test,was carried out for NC gels and M-NCs. The safety of NC gels and M-NCs was confirmed in all tests. Also, the interaction of living tissue with NC gel was investigated in vivo by implantation in live goats; neither inflammation nor concrescence occurred around the NC gels. Furthermore, it was found that both N-NC gels consisting of poly(N-isopropylacrylamide(PNIPA/clay network and M-NCs consisting of poly(2-methoxyethyacrylate(PMEA/clay network show characteristic cell culture and subsequent cell detachment on their surfaces, although it was almost impossible to culture cells on conventional, chemically-crosslinked PNIPA hydrogels and chemically crossslinked PMEA, regardless of their crosslinker concentration. Various kinds of cells, such ashumanhepatoma cells (HepG2, normal human dermal fibroblast (NHDF, and human umbilical vein endothelial cells (HUVEC, could be cultured to be confluent on the surfaces of N
-
Recombinant Protein Production and Insect Cell Culture and Process
Spaulding, Glenn F. (Inventor); Goodwin, Thomas J. (Inventor); OConnor, Kim C. (Inventor); Francis, Karen M. (Inventor); Andrews, Angela D. (Inventor); Prewett, Tracey L. (Inventor)
1997-01-01
A process has been developed for recombinant production of selected polypeptides using transformed insect cells cultured in a horizontally rotating culture vessel modulated to create low shear conditions. A metabolically transformed insect cell line is produced using the culture procedure regardless of genetic transformation. The recombinant polypeptide can be produced by an alternative process using virtually infected or stably transformed insect cells containing a gene encoding the described polypeptide. The insect cells can also be a host for viral production.
-
Metabolite profiling of microfluidic cell culture conditions for droplet based screening
DEFF Research Database (Denmark)
Björk, Sara M.; Sjoström, Staffan L.; Svahn, Helene Andersson
2015-01-01
We investigate the impact of droplet culture conditions on cell metabolic state by determining key metabolite concentrations in S. cerevisiae cultures in different microfluidic droplet culture formats. Control of culture conditions is critical for single cell/clone screening in droplets......, such as directed evolution of yeast, as cell metabolic state directly affects production yields from cell factories. Here, we analyze glucose, pyruvate, ethanol, and glycerol, central metabolites in yeast glucose dissimilation to establish culture formats for screening of respiring as well as fermenting yeast...... limited cultures, whereas the metabolite profiles of cells cultured in the alternative wide tube droplet incubation format resemble those from aerobic culture. Furthermore, we demonstrate retained droplet stability and size in the new better oxygenated droplet incubation format....
-
Strachan, Samantha L.
The underachievement of African American students in science has been a persistent problem in science education. The achievement patterns of African American students indicate that researchers must take a closer look at the types of practices that are being used to meet these students' needs in science classrooms. Determining why science teachers decide to employ certain practices in their classrooms begins with a careful examination of teachers' beliefs as well as their instructional approaches. The purpose of this study was to explore four urban high school science teachers' beliefs about their African American students' learning needs and to investigate how these teachers go about addressing students' needs in science classrooms. This research study also explored the extent to which teachers' practices aligned with the nine dimensions of an established cultural instructional theory, namely the Black Cultural Ethos. Qualitative research methods were employed to gather data from the four teachers. Artifact data were collected from the teachers and they were interviewed and observed. Believing that their students had academic-related needs as well as needs tied to their learning preferences, the four science teachers employed a variety of instructional strategies to meet their students where they were in learning. Overall, the instructional strategies that the teachers employed to meet their students' needs aligned with five of the nine tenets of the Black Cultural Ethos theory.
-
A Cell Culture Approach to Optimized Human Corneal Endothelial Cell Function
Bartakova, Alena; Kuzmenko, Olga; Alvarez-Delfin, Karen; Kunzevitzky, Noelia J.; Goldberg, Jeffrey L.
2018-01-01
Purpose Cell-based therapies to replace corneal endothelium depend on culture methods to optimize human corneal endothelial cell (HCEC) function and minimize endothelial-mesenchymal transition (EnMT). Here we explore contribution of low-mitogenic media on stabilization of phenotypes in vitro that mimic those of HCECs in vivo. Methods HCECs were isolated from cadaveric donor corneas and expanded in vitro, comparing continuous presence of exogenous growth factors (“proliferative media”) to media without those factors (“stabilizing media”). Identity based on canonical morphology and expression of surface marker CD56, and function based on formation of tight junction barriers measured by trans-endothelial electrical resistance assays (TEER) were assessed. Results Primary HCECs cultured in proliferative media underwent EnMT after three to four passages, becoming increasingly fibroblastic. Stabilizing the cells before each passage by switching them to a media low in mitogenic growth factors and serum preserved canonical morphology and yielded a higher number of cells. HCECs cultured in stabilizing media increased both expression of the identity marker CD56 and also tight junction monolayer integrity compared to cells cultured without stabilization. Conclusions HCECs isolated from donor corneas and expanded in vitro with a low-mitogenic media stabilizing step before each passage demonstrate more canonical structural and functional features and defer EnMT, increasing the number of passages and total canonical cell yield. This approach may facilitate development of HCEC-based cell therapies. PMID:29625488
-
Sponge cell culture? A molecular identification method for sponge cells
Sipkema, D.; Heilig, G.H.J.; Akkermans, A.D.L.; Osinga, R.; Tramper, J.; Wijffels, R.H.
2003-01-01
Dissociated sponge cells are easily confused with unicellular organisms. This has been an obstacle in the development of sponge-cell lines. We developed a molecular detection method to identify cells of the sponge Dysidea avara in dissociated cell cultures. The 18S ribosomal RNA gene from a Dysidea
-
Akahane, M; Shimizu, T; Kira, T; Onishi, T; Uchihara, Y; Imamura, T; Tanaka, Y
2016-11-01
To assess the structure and extracellular matrix molecule expression of osteogenic cell sheets created via culture in medium with both dexamethasone (Dex) and ascorbic acid phosphate (AscP) compared either Dex or AscP alone. Osteogenic cell sheets were prepared by culturing rat bone marrow stromal cells in a minimal essential medium (MEM), MEM with AscP, MEM with Dex, and MEM with Dex and AscP (Dex/AscP). The cell number and messenger (m)RNA expression were assessed in vitro, and the appearance of the cell sheets was observed after mechanical retrieval using a scraper. β-tricalcium phosphate (β-TCP) was then wrapped with the cell sheets from the four different groups and subcutaneously implanted into rats. After mechanical retrieval, the osteogenic cell sheets from the MEM, MEM with AscP, and MEM with Dex groups appeared to be fragmented or incomplete structures. The cell sheets cultured with Dex/AscP remained intact after mechanical retrieval, without any identifiable tears. Culture with Dex/AscP increased the mRNA and protein expression of extracellular matrix proteins and cell number compared with those of the other three groups. More bridging bone formation was observed after transplantation of the β-TCP scaffold wrapped with cell sheets cultured with Dex/AscP, than in the other groups. These results suggest that culture with Dex/AscP improves the mechanical integrity of the osteogenic cell sheets, allowing retrieval of the confluent cells in a single cell sheet structure. This method may be beneficial when applied in cases of difficult tissue reconstruction, such as nonunion, bone defects, and osteonecrosis.Cite this article: M. Akahane, T. Shimizu, T. Kira, T. Onishi, Y. Uchihara, T. Imamura, Y. Tanaka. Culturing bone marrow cells with dexamethasone and ascorbic acid improves osteogenic cell sheet structure. Bone Joint Res 2016;5:569-576. DOI: 10.1302/2046-3758.511.BJR-2016-0013.R1. © 2016 Akahane et al.
-
Yoshimitsu, Ryosuke; Hattori, Koji; Sugiura, Shinji; Kondo, Yuki; Yamada, Rotaro; Tachikawa, Saoko; Satoh, Taku; Kurisaki, Akira; Ohnuma, Kiyoshi; Asashima, Makoto; Kanamori, Toshiyuki
2014-05-01
Human induced pluripotent stem cells (hiPSCs) are a promising cell source for drug screening. For this application, self-renewal or differentiation of the cells is required, and undefined factors in the culture conditions are not desirable. Microfluidic perfusion culture allows the production of small volume cultures with precisely controlled microenvironments, and is applicable to high-throughput cellular environment screening. Here, we developed a microfluidic perfusion culture system for hiPSCs that uses a microchamber array chip under defined extracellular matrix (ECM) and culture medium conditions. By screening various ECMs we determined that fibronectin and laminin are appropriate for microfluidic devices made out of the most popular material, polydimethylsiloxane (PDMS). We found that the growth rate of hiPSCs under pressure-driven perfusion culture conditions was higher than under static culture conditions in the microchamber array. We applied our new system to self-renewal and differentiation cultures of hiPSCs, and immunocytochemical analysis showed that the state of the hiPSCs was successfully controlled. The effects of three antitumor drugs on hiPSCs were comparable between microchamber array and 96-well plates. We believe that our system will be a platform technology for future large-scale screening of fully defined conditions for differentiation cultures on integrated microfluidic devices. © 2013 Wiley Periodicals, Inc.
-
Quantitative volumetric Raman imaging of three dimensional cell cultures
Kallepitis, Charalambos; Bergholt, Mads S.; Mazo, Manuel M.; Leonardo, Vincent; Skaalure, Stacey C.; Maynard, Stephanie A.; Stevens, Molly M.
2017-03-01
The ability to simultaneously image multiple biomolecules in biologically relevant three-dimensional (3D) cell culture environments would contribute greatly to the understanding of complex cellular mechanisms and cell-material interactions. Here, we present a computational framework for label-free quantitative volumetric Raman imaging (qVRI). We apply qVRI to a selection of biological systems: human pluripotent stem cells with their cardiac derivatives, monocytes and monocyte-derived macrophages in conventional cell culture systems and mesenchymal stem cells inside biomimetic hydrogels that supplied a 3D cell culture environment. We demonstrate visualization and quantification of fine details in cell shape, cytoplasm, nucleus, lipid bodies and cytoskeletal structures in 3D with unprecedented biomolecular specificity for vibrational microspectroscopy.
-
Building a Science Communication Culture: One Agency's Approach
DeWitt, S.; Tenenbaum, L. F.; Betz, L.
2014-12-01
Science communication does not have to be a solitary practice. And yet, many scientists go about it alone and with little support from their peers and organizations. To strengthen community and build support for science communicators, NASA designed a training course aimed at two goals: 1) to develop individual scientists' communication skills, and 2) to begin to build a science communication culture at the agency. NASA offered a pilot version of this training course in 2014: the agency's first multidisciplinary face-to-face learning experience for science communicators. Twenty-six Earth, space and life scientists from ten field centers came together for three days of learning. They took part in fundamental skill-building exercises, individual development planning, and high-impact team projects. This presentation will describe the course design and learning objectives, the experience of the participants, and the evaluation results that will inform future offerings of communication training for NASA scientists and others.
-
Some Lessons From a Symposium on Cultural Psychological Science.
Sternberg, Robert J
2017-09-01
In this concluding essay, I summarize some of the main points of each of the contributors and attempt to highlight their importance for psychological science and for everyday life. I bring in some examples of research from my own research group over the years that reinforce many of the conclusions reached by the contributors. The purpose of this symposium on cultural psychological science is, we hope, to teach some lessons that could not easily be learned except through cultural research. My goal in this final essay is to consider what I believe to be a primary lesson of each contribution. I attempt to illustrate the considerable relevance of each of these contributions to contemporary society. The views expressed here are solely my own, and of course readers may find much to disagree with; hopefully, they will find some things to agree with as well!
-
Chen, Yi-Dao; Huang, Shiang-Fu; Wang, Hung-Ming
2015-01-01
To precisely and faithfully perform cell-based drug chemosensitivity assays, a well-defined and biologically relevant culture condition is required. For the former, a perfusion microbioreactor system capable of providing a stable culture condition was adopted. For the latter, however, little is known about the impact of culture models on the physiology and chemosensitivity assay results of primary oral cavity cancer cells. To address the issues, experiments were performed. Results showed that minor environmental pH change could significantly affect the metabolic activity of cells, demonstrating the importance of stable culture condition for such assays. Moreover, the culture models could also significantly influence the metabolic activity and proliferation of cells. Furthermore, the choice of culture models might lead to different outcomes of chemosensitivity assays. Compared with the similar test based on tumor-level assays, the spheroid model could overestimate the drug resistance of cells to cisplatin, whereas the 2D and 3D culture models might overestimate the chemosensitivity of cells to such anticancer drug. In this study, the 3D culture models with same cell density as that in tumor samples showed comparable chemosensitivity assay results as the tumor-level assays. Overall, this study has provided some fundamental information for establishing a precise and faithful drug chemosensitivity assay. PMID:25654105
-
Fekete, Natalie; Béland, Ariane V; Campbell, Katie; Clark, Sarah L; Hoesli, Corinne A
2018-04-19
In recent years, cell-based therapies targeting the immune system have emerged as promising strategies for cancer treatment. This review summarizes manufacturing challenges related to production of antigen presenting cells as a patient-tailored cancer therapy. Understanding cell-material interactions is essential because in vitro cell culture manipulations to obtain mature antigen-producing cells can significantly alter their in vivo performance. Traditional antigen-producing cell culture protocols often rely on cell adhesion to surface-treated hydrophilic polystyrene flasks. More recent commercial and investigational cancer immunotherapy products were manufactured using suspension cell culture in closed hydrophobic fluoropolymer bags. The shift to closed cell culture systems can decrease risks of contamination by individual operators, as well as facilitate scale-up and automation. Selecting closed cell culture bags over traditional open culture systems entails different handling procedures and processing controls, which can affect product quality. Changes in culture vessels also entail changes in vessel materials and geometry, which may alter the cell microenvironment and resulting cell fate decisions. Strategically designed culture systems will pave the way for the generation of more sophisticated and highly potent cell-based cancer vaccines. As an increasing number of cell-based therapies enter the clinic, the selection of appropriate cell culture vessels and materials becomes a critical consideration that can impact the therapeutic efficacy of the product, and hence clinical outcomes and patient quality of life. © 2018 The Authors Transfusion published by Wiley Periodicals, Inc. on behalf of AABB.
-
Three-dimensional hydrogel cell culture systems for modeling neural tissue
Frampton, John
Two-dimensional (2-D) neural cell culture systems have served as physiological models for understanding the cellular and molecular events that underlie responses to physical and chemical stimuli, control sensory and motor function, and lead to the development of neurological diseases. However, the development of three-dimensional (3-D) cell culture systems will be essential for the advancement of experimental research in a variety of fields including tissue engineering, chemical transport and delivery, cell growth, and cell-cell communication. In 3-D cell culture, cells are provided with an environment similar to tissue, in which they are surrounded on all sides by other cells, structural molecules and adhesion ligands. Cells grown in 3-D culture systems display morphologies and functions more similar to those observed in vivo, and can be cultured in such a way as to recapitulate the structural organization and biological properties of tissue. This thesis describes a hydrogel-based culture system, capable of supporting the growth and function of several neural cell types in 3-D. Alginate hydrogels were characterized in terms of their biomechanical and biochemical properties and were functionalized by covalent attachment of whole proteins and peptide epitopes. Methods were developed for rapid cross-linking of alginate hydrogels, thus permitting the incorporation of cells into 3-D scaffolds without adversely affecting cell viability or function. A variety of neural cell types were tested including astrocytes, microglia, and neurons. Cells remained viable and functional for longer than two weeks in culture and displayed process outgrowth in 3-D. Cell constructs were created that varied in cell density, type and organization, providing experimental flexibility for studying cell interactions and behavior. In one set of experiments, 3-D glial-endothelial cell co-cultures were used to model blood-brain barrier (BBB) structure and function. This co-culture system was
-
Azizollah, Arbabisarjou; Abolghasem, Farhang; Amin, Dadgar Mohammad
2016-01-01
Background and Objective: Organizations effort is to achieve a common goal. There are many constructs needed for organizations. Organizational culture and organizational commitment are special concepts in management. The objective of the current research is to study the relationship between organizational culture and organizational commitment among the personnel of Zahedan University of Medical Sciences. Materials and Methods: This is a descriptive- correlational study. The statistical population was whole tenured staff of Zahedan University of Medical Sciences that worked for this organization in 2012-2013. Random sampling method was used and 165 samples were chosen. Two standardized questionnaires of the organizational culture (Schein, 1984) and organizational commitment (Meyer & Allen, 2002) were applied. The face and construct validity of the questionnaires were approved by the lecturers of Management and experts. Reliability of questionnaires of the organizational culture and organizational commitment were 0.89 and 0.88 respectively, by Cronbach’s Alpha coefficient. All statistical calculations performed using Statistical Package for the Social Sciences version 21.0 (SPSS Inc., Chicago, IL, USA). The level of significance was set at Porganizational culture and organizational commitment (P value=0.027). Also, the results showed that there was a significant relation between organizational culture and affective commitment (P-value=0.009), organizational culture and continuance commitment (P-value=0.009), and organizational culture and normative commitment (P-value=0.009). PMID:26925884
-
Azizollah, Arbabisarjou; Abolghasem, Farhang; Mohammad Amin, Dadgar
2015-12-14
Organizations effort is to achieve a common goal. There are many constructs needed for organizations. Organizational culture and organizational commitment are special concepts in management. The objective of the current research is to study the relationship between organizational culture and organizational commitment among the personnel of Zahedan University of Medical Sciences. This is a descriptive- correlational study. The statistical population was whole tenured staff of Zahedan University of Medical Sciences that worked for this organization in 2012-2013. Random sampling method was used and 165 samples were chosen. Two standardized questionnaires of the organizational culture (Schein, 1984) and organizational commitment (Meyer & Allen, 2002) were applied. The face and construct validity of the questionnaires were approved by the lecturers of Management and experts. Reliability of questionnaires of the organizational culture and organizational commitment were 0.89 and 0.88 respectively, by Cronbach's Alpha coefficient. All statistical calculations performed using Statistical Package for the Social Sciences version 21.0 (SPSS Inc., Chicago, IL, USA). The level of significance was set at Porganizational culture and organizational commitment (P value=0.027). Also, the results showed that there was a significant relation between organizational culture and affective commitment (P-value=0.009), organizational culture and continuance commitment (P-value=0.009), and organizational culture and normative commitment (P-value=0.009).
-
International Nuclear Information System (INIS)
Tokita, N.; Carpenter, S.G.; Raju, M.R.
1984-01-01
Cell-cycle distributions were measured by flow cytometry for Chinese hamster (CHO) cells cultured continuously under hypoxic conditions. DNA histograms showed an accumulation of cells in the early S phase followed by a traverse delay through the S phase, and a G 2 block. During hypoxic culturing, cell viability decreased rapidly to less than 0.1% at 120 h. Radiation responses for cells cultured under these conditions showed an extreme radioresistance at 72 h. Results suggest that hypoxia induces a condition similar to cell synchrony which itself changes the radioresistance of hypoxic cells. (author)
-
Kaji, Hirokazu; Camci-Unal, Gulden; Langer, Robert; Khademhosseini, Ali
2011-03-01
Inside the body, cells lie in direct contact or in close proximity to other cell types in a tightly controlled architecture that often regulates the resulting tissue function. Therefore, tissue engineering constructs that aim to reproduce the architecture and the geometry of tissues will benefit from methods of controlling cell-cell interactions with microscale resolution. We discuss the use of microfabrication technologies for generating patterned co-cultures. In addition, we categorize patterned co-culture systems by cell type and discuss the implications of regulating cell-cell interactions in the resulting biological function of the tissues. Patterned co-cultures are a useful tool for fabricating tissue engineered constructs and for studying cell-cell interactions in vitro, because they can be used to control the degree of homotypic and heterotypic cell-cell contact. In addition, this approach can be manipulated to elucidate important factors involved in cell-matrix interactions. Patterned co-culture strategies hold significant potential to develop biomimetic structures for tissue engineering. It is expected that they would create opportunities to develop artificial tissues in the future. This article is part of a Special Issue entitled Nanotechnologies - Emerging Applications in Biomedicine. 2010 Elsevier B.V. All rights reserved.
-
Gebhard, C; Gabriel, C; Walter, I
2016-06-01
Spheroid cell culture emerges as powerful in vitro tool for experimental tumour research. In this study, we established a scaffold-free three-dimensional spheroid system built from canine osteosarcoma (OS) cells (D17). Spheroids (7, 14 and 19 days of cultivation) and monolayer cultures (2 and 7 days of cultivation) were evaluated and compared on light and electron microscopy. Monolayer and spheroid cultures were tested for vimentin, cytokeratin, alkaline phosphatase, osteocalcin and collagen I by means of immunohistochemistry. The spheroid cell culture exhibited a distinct network of collagen I in particular after 19-day cultivation, whereas in monolayer cultures, collagen I was arranged as a lamellar basal structure. Necrotic centres of large spheroids, as observed in 14- and 19-day cultures, were characterized by significant amounts of osteocalcin. Proliferative activity as determined by Ki-67 immunoreactivity showed an even distribution in two-dimensional cultures. In spheroids, proliferation was predominating in the peripheral areas. Metastasis-associated markers ezrin and S100A4 were shown to be continuously expressed in monolayer and spheroid cultures. We conclude that the scaffold-free spheroid system from canine OS cells has the ability to mimic the architecture of the in vivo tumour, in particular cell-cell and cell-matrix interactions. © 2015 The Authors. Anatomia, Histologia, Embryologia Published by Blackwell Verlag GmbH.
-
A microwell cell culture platform for the aggregation of pancreatic β-cells.
Bernard, Abigail B; Lin, Chien-Chi; Anseth, Kristi S
2012-08-01
Cell-cell contact between pancreatic β-cells is important for maintaining survival and normal insulin secretion. Various techniques have been developed to promote cell-cell contact between β-cells, but a simple yet robust method that affords precise control over three-dimensional (3D) β-cell cluster size has not been demonstrated. To address this need, we developed a poly(ethylene glycol) (PEG) hydrogel microwell platform using photolithography. This microwell cell-culture platform promotes the formation of 3D β-cell aggregates of defined sizes from 25 to 210 μm in diameter. Using this platform, mouse insulinoma 6 (MIN6) β-cells formed aggregates with cell-cell adherin junctions. These naturally formed cell aggregates with controllable sizes can be removed from the microwells for macroencapsulation, implantation, or other biological assays. When removed and subsequently encapsulated in PEG hydrogels, the aggregated cell clusters demonstrated improved cellular viability (>90%) over 7 days in culture, while the β-cells encapsulated as single cells maintained only 20% viability. Aggregated MIN6 cells also exhibited more than fourfold higher insulin secretion in response to a glucose challenge compared with encapsulated single β-cells. Further, the cell aggregates stained positively for E-cadherin, indicative of the formation of cell junctions. Using this hydrogel microwell cell-culture method, viable and functional β-cell aggregates of specific sizes were created, providing a platform from which other biologically relevant questions may be answered.
-
Quantitative volumetric Raman imaging of three dimensional cell cultures
Kallepitis, Charalambos
2017-03-22
The ability to simultaneously image multiple biomolecules in biologically relevant three-dimensional (3D) cell culture environments would contribute greatly to the understanding of complex cellular mechanisms and cell–material interactions. Here, we present a computational framework for label-free quantitative volumetric Raman imaging (qVRI). We apply qVRI to a selection of biological systems: human pluripotent stem cells with their cardiac derivatives, monocytes and monocyte-derived macrophages in conventional cell culture systems and mesenchymal stem cells inside biomimetic hydrogels that supplied a 3D cell culture environment. We demonstrate visualization and quantification of fine details in cell shape, cytoplasm, nucleus, lipid bodies and cytoskeletal structures in 3D with unprecedented biomolecular specificity for vibrational microspectroscopy.
-
Cell division requirement for activation of murine leukemia virus in cell culture by irradiation
International Nuclear Information System (INIS)
Otten, J.A.; Quarles, J.M.; Tennant, R.W.
1976-01-01
Actively dividing cultures of AKR mouse cells were exposed to relatively low dose-rates of γ radiation and tested for activation of endogenous leukemia viruses. Efficient and reproducible induction of virus was obtained with actively dividing cells, but cultures deprived of serum to inhibit cell division before and during γ irradiation were not activated, even when medium with serum was added immediately after irradiation. These results show that cell division was required for virus induction but that a stable intermediate similar to the state induced by halogenated pyrimidines was not formed. In actively dividing AKR cell cultures, virus activation appeared to be proportional to the dose of γ radiation; the estimated frequency of activation was 1-8 x 10 - 5 per exposed cell and the efficiency of activation was approximately 0.012 inductions per cell per rad. Other normal primary and established mouse cell cultures tested were not activated by γ radiation. The requirement of cell division for radiation and chemical activation may reflect some common mechanism for initiation of virus expression
-
Bradley, Jillian H.; Stein, Rachel; Randolph, Brad; Molina, Emily; Arnold, Jennifer P.; Gregg, Randal K.
2017-11-01
Immune impairment mediated by microgravity threatens the success of space exploration requiring long-duration spaceflight. The cells of most concern, T lymphocytes, coordinate the host response against microbial and cancerous challenges leading to elimination and long-term protection. T cells are activated upon recognition of specific microbial peptides bound on the surface of antigen presenting cells, such as dendritic cells (DC). Subsequently, this engagement results in T cell proliferation and differentiation into effector T cells driven by autocrine interleukin-2 (IL-2) and other cytokines. Finally, the effector T cells acquire the weaponry needed to destroy microbial invaders and tumors. Studies conducted on T cells during spaceflight, or using Earth-based culture systems, have shown reduced production of cytokines, proliferation and effector functions as compared to controls. This may account for the cases of viral reactivation events and opportunistic infections associated with astronauts of numerous missions. This work has largely been based upon the outcome of T cell activation by stimulatory factors that target select T cell signaling pathways rather than the complex, signaling events related to the natural process of antigen presentation by DC. This study tested the response of an ovalbumin peptide-specific T cell line, OT-II TCH, to activation by DC when the T cells were cultured 24-120 h in a simulated microgravity (SMG) environment generated by a rotary cell culture system. Following 72 h culture of T cells in SMG (SMG-T) or control static (Static-T) conditions, IL-2 production by the T cells was reduced in SMG-T cells compared to Static-T cells upon stimulation by phorbol 12-myristate 13-acetate (PMA) and ionomycin. However, when the SMG-T cells were stimulated with DC and peptide, IL-2 was significantly increased compared to Static-T cells. Such enhanced IL-2 production by SMG-T cells peaked at 72 h SMG culture time and decreased thereafter. When
-
Tumor necrosis factor (cachetin) decreases adipose cell differentiation in primary cell culture
International Nuclear Information System (INIS)
Martin, R.J.; Jones, D.D.; Jewell, D.E.; Hausman, G.J.
1986-01-01
Cachetin has been shown to effect gene product expression in the established adipose cell line 3T3-L1. Expression of messenger RNA for lipoprotein lipase is suppressed in cultured adipocytes. The purpose of this study was to determine the effect of Cachetin on adipose cell differentiation in primary cell culture. Stromalvascular cells obtained from the inguinal fat pad of 4-5 week old Sprague-Dawley rats were grown in culture for two weeks. During the proliferative growth phase all cells were grown on the same medium and labelled with 3 H-thymidine. Cachetin treatment (10 -6 to 10 -10 M) was initiated on day 5, the initial phase of preadipocyte differentiation. Adipocytes and stromal cells were separated using density gradient, and 3 H-thymidine was determined for both cell types. Thymidine incorporation into adipose cells was decreased maximally (∼ 50%) at 10 -10 M. Stromalvascular cells were not influenced at any of the doses tested. Adipose cell lipid content as indicated by oil red-O staining was decreased by Cachetin. Esterase staining by adipose cells treated with Cachetin was increased indicating an increase in intracellular lipase. These studies show that Cachetin has specific effects on primary adipose cell differentiation
-
Feeding Frequency Affects Cultured Rat Pituitary Cells in Low Gravity
Hymer, W. C.; Grindeland, R. E.; Salada, T.; Cenci, R.; Krishnan, K.; Mukai, C.; Nagaoka, S.
1996-01-01
In this report, we describe the results of a rat pituitary cell culture experiment done on STS-65 in which the effect of cell feeding on the release of the six anterior pituitary hormones was studied. We found complex microgravity related interactions between the frequency of cell feeding and the quantity and quality (i.e. biological activity) of some of the six hormones released in flight. Analyses of growth hormone (GH) released from cells into culture media on different mission days using gel filtration and ion exchange chromatography yielded qualitatively similar results between ground and flight samples. Lack of cell feeding resulted in extensive cell clumping in flight (but not ground) cultures. Vigorous fibroblast growth occurred in both ground and flight cultures fed 4 times. These results are interpreted within the context of autocrine and or paracrine feedback interactions. Finally the payload specialist successfully prepared a fresh trypsin solution in microgravity, detached the cells from their surface and reinserted them back into the culture chamber. These cells reattached and continued to release hormone in microgravity. In summary, this experiment shows that pituitary cells are microgravity sensitive and that coupled operations routinely associated with laboratory cel1 culture can also be accomplished in low gravity.
-
Hằng, Ngô Vũ Thu; Meijer, Marijn Roland; Bulte, Astrid M. W.; Pilot, Albert
2017-09-01
The implementation of social constructivist approaches to learning science in primary education in Vietnamese culture as an example of Confucian heritage culture remains challenging and problematic. This theoretical paper focuses on the initial phase of a design-based research approach; that is, the description of the design of a formal, written curriculum for primary science education in which features of social constructivist approaches to learning are synthesized with essential aspects of Vietnamese culture. The written design comprises learning aims, a framework that is the synthesis of learning functions, learning settings and educational expectations for learning phases, and exemplary curriculum units. Learning aims are formulated to comprehensively develop scientific knowledge, skills, and attitudes toward science for primary students. Derived from these learning aims, the designed framework consists of four learning phases respectively labeled as Engagement, Experience, Exchange, and Follow-up. The designed framework refers to knowledge of the "nature of science" education and characteristics of Vietnamese culture as an example of Confucian heritage culture. The curriculum design aims to serve as an educational product that addresses previously analyzed problems of primary science education in the Vietnamese culture in a globalizing world.
-
Energy Technology Data Exchange (ETDEWEB)
Ortiz de Montellano, B.
1996-11-14
As planned a letter was sent out to 17 teachers who had participated in a Summer 1994 workshop on ``Culturally Relevant Science for Hispanics`` at Michigan State. These teachers were supposed to have spent the intervening time developing lesson plans and curricula. The letter requested a report of any activities undertaken and copies of lesson plans and materials developed by February 1996 with a stipend of $400 for satisfactory reports. It was a disappointment to only get 9 responses and not all of them demonstrating a satisfactory level of activity. Diana Marinez, Dean of Science at Texas A and M University, Corpus Christi, who is the other developer of this curriculum and the author reviewed the submitted materials and chose those showing the most promise to be invited to participate in the Summer Writing Workshop. Spring of 1996 and particularly in May--June, the author wrote a partial first draft of a companion volume for the teacher`s manual which would provide a rationale for doing culturally relevant science, present the cultural and the scientific background that teachers would need in order to be able to teach. One of the goals of this curriculum is that it should be off-the-shelf ready to teach and that teachers would not have to do extra research to encourage its adoption. The outline of the book is appendix 1. The Writing Workshop was held at Texas A and M University, Corpus Christi from July 14 to July 27, 1996. Participating teachers chose topics that they were interested in developing and wrote first drafts. These were distributed to all participants and critiqued by the workshop directors before being rewritten. Some teachers were more productive than others depending on their science background. In total an impressive number of lesson plans were written. These lesson plans are listed in Appendix 3. Appendix 4 is a sample lesson. Work still needs to be done on both the source book and the teachers` manual.
-
Harnessing Science and Technology for preservation and conservation of cultural heritage in Malaysia
International Nuclear Information System (INIS)
Adi Taha
2005-04-01
Malaysia's heritage is extraordinarily rich. Heritage links people, places and things from our history to the present and to the future. Department of Muzeums and Antiquities work diligently at collecting and preserving the artifacts, written records, oral traditions, special places and lands that make up the Malaysia's history. Over the years our concept of cultural heritage and its role as a central part of the experience of our communities has expanded from a focus on objects and monuments to include our social structures, ways of life, beliefs and systems of knowledge. We seek answers in our attempts to promote the understanding and unity among people that have made our country a nation regardless of ethnic origins and religious affiliations, and to prolong the life essence of our rich heritage. We found a simple but yet, a meaningful answer; Harnessing Science and Technology for Preservation and Conservation of Cultural Heritage in Malaysia. Conservation has gained an increasing importance world over, as there is greater awareness and a sense of urgency about the need to conserve and preserve cultural heritages. Recent years are witnessing unprecedented growth in various fields of science and technology in Malaysia, such as materials technology, medical sciences, biotechnology, information and communications technology. Whichever perspective is used, it is clear that science forms an integral part of Malaysia's culture, in the past as well as now. Fulfilling a vital function as a carrier of knowledge and methodology, sciences places on our shoulders a strong obligation towards future generations. As Malaysians, we have been formed by our cultural heritage. Clearly, we must protect that heritage and continue to enrich and develop it, incorporating new knowledge, new insights, new ideas and new experience. (Author)
-
Impact of Chinese Culture on Pre-service Science Teachers' Views of the Nature of Science
Wan, Dongsheng; Zhang, Hongshia; Wei, Bing
2018-04-01
This study examines Chinese pre-service teachers' (N = 30) views on the nature of science (NOS) and how Chinese culture influences their views. Participants were from two teachers' universities in eastern China. As an exploratory and interpretive study, a scenario-based interview approach was adopted. The results indicated that the participants held unique views about the five key aspects of NOS. Many participants have alternative and contemporary views of NOS, but few possess classical views. In fact, teachers adopted features of the Confucian Doctrine of the Mean either consciously or unconsciously to account for their views of NOS. This research reflects that the Doctrine of the Mean affected Chinese teachers' views of NOS, making them rather deficient in their understandings of classical NOS. Based on empirical data, it is argued that science teacher training in China should focus on the content and objectives of classical NOS, rather than just teaching contemporary views of NOS. Taking Chinese culture into consideration, science teacher education in China cannot entirely import the strategies of teaching the classical views of NOS from the developed world, but should develop, design and contextualize local strategies that are suitable for the training of Chinese science teachers. Some issues for further investigation of learners' views of NOS in non-Western contexts are suggested as implications from this study.
-
Cultural psychology as a bridge between anthropology and cognitive science.
Fryberg, Stephanie A
2012-07-01
The theory and methods of cultural psychology begin with the assumption that psychological processes are socioculturally and historically grounded. As such, they offer a new approach for understanding the diversity of human functioning because they (a) question the presumed neutrality of the majority group perspective; (b) take the target's point-of-view (i.e., what it means to be a person in a particular context); (c) assume that there is more than one viable way of being a competent or effective person; and (d) provide a road map for understanding and reducing social inequities. As illustrated in this essay, a cultural psychological approach provides a bridge between anthropology and the cognitive sciences, and in so doing it offers an alternative set of explanations and interventions for group differences. Copyright © 2012 Cognitive Science Society, Inc.
-
Jin, Liting; Qu, Ying; Gomez, Liliana J; Chung, Stacey; Han, Bingchen; Gao, Bowen; Yue, Yong; Gong, Yiping; Liu, Xuefeng; Amersi, Farin; Dang, Catherine; Giuliano, Armando E; Cui, Xiaojiang
2018-02-20
Conditional reprogramming methods allow for the inexhaustible in vitro proliferation of primary epithelial cells from human tissue specimens. This methodology has the potential to enhance the utility of primary cell culture as a model for mammary gland research. However, few studies have systematically characterized this method in generating in vitro normal human mammary epithelial cell models. We show that cells derived from fresh normal breast tissues can be propagated and exhibit heterogeneous morphologic features. The cultures are composed of CK18, desmoglein 3, and CK19-positive luminal cells and vimentin, p63, and CK14-positive myoepithelial cells, suggesting the maintenance of in vivo heterogeneity. In addition, the cultures contain subpopulations with different CD49f and EpCAM expression profiles. When grown in 3D conditions, cells self-organize into distinct structures that express either luminal or basal cell markers. Among these structures, CK8-positive cells enclosing a lumen are capable of differentiation into milk-producing cells in the presence of lactogenic stimulus. Furthermore, our short-term cultures retain the expression of ERα, as well as its ability to respond to estrogen stimulation. We have investigated conditionally reprogrammed normal epithelial cells in terms of cell type heterogeneity, cellular marker expression, and structural arrangement in two-dimensional (2D) and three-dimensional (3D) systems. The conditional reprogramming methodology allows generation of a heterogeneous culture from normal human mammary tissue in vitro . We believe that this cell culture model will provide a valuable tool to study mammary cell function and malignant transformation.
-
Pushing the boundaries of cultural congruence pedagogy in science education towards a third space
Quigley, Cassie
2011-09-01
This review explores Meyers and Crawford's "Teaching science as a cultural way of knowing: Merging authentic inquiry, nature of science, and multicultural strategies" by examining how they combine the use of inquiry-based science instruction with multicultural strategies. In this conversation, I point to the need of specific discourse strategies to help teachers and students create hybrid spaces to push the boundaries of cultural congruence as described in this article. These strategies include a reflective component to the explicit instruction that encourages an integration of home and science discourses. My response to this work expands on their use of multicultural strategies to push toward a congruent Third space that asks not only what happens to the students who do not participate in science, but also what happens to science when a diverse group of people does not participate?
-
Gabashvili, A N; Baklaushev, V P; Grinenko, N F; Mel'nikov, P A; Cherepanov, S A; Levinsky, A B; Chehonin, V P
2016-02-01
The tumor-suppressive effect of rat mesenchymal stem cells against low-differentiated rat C6 glioma cells during their direct and indirect co-culturing and during culturing of C6 glioma cells in the medium conditioned by mesenchymal stem cells was studied in an in vitro experiment. The most pronounced antitumor activity of mesenchymal stem cells was observed during direct co-culturing with C6 glioma cells. The number of live C6 glioma cells during indirect co-culturing and during culturing in conditioned medium was slightly higher than during direct co-culturing, but significantly differed from the control (C6 glioma cells cultured in medium conditioned by C6 glioma cells). The cytotoxic effect of medium conditioned by mesenchymal stem cells was not related to medium depletion by glioma cells during their growth. The medium conditioned by other "non-stem" cells (rat astrocytes and fibroblasts) produced no tumor-suppressive effect. Rat mesenchymal stem cells, similar to rat C6 glioma cells express connexin 43, the main astroglial gap junction protein. During co-culturing, mesenchymal stem cells and glioma C6 cells formed functionally active gap junctions. Gap junction blockade with connexon inhibitor carbenoxolone attenuated the antitumor effect observed during direct co-culturing of C6 glioma cells and mesenchymal stem cells to the level produced by conditioned medium. Cell-cell signaling mediated by gap junctions can be a mechanism of the tumor-suppressive effect of mesenchymal stem cells against C6 glioma cells. This phenomenon can be used for the development of new methods of cell therapy for high-grade malignant gliomas.
-
Advances in tissue engineering through stem cell-based co-culture.
Paschos, Nikolaos K; Brown, Wendy E; Eswaramoorthy, Rajalakshmanan; Hu, Jerry C; Athanasiou, Kyriacos A
2015-05-01
Stem cells are the future in tissue engineering and regeneration. In a co-culture, stem cells not only provide a target cell source with multipotent differentiation capacity, but can also act as assisting cells that promote tissue homeostasis, metabolism, growth and repair. Their incorporation into co-culture systems seems to be important in the creation of complex tissues or organs. In this review, critical aspects of stem cell use in co-culture systems are discussed. Direct and indirect co-culture methodologies used in tissue engineering are described, along with various characteristics of cellular interactions in these systems. Direct cell-cell contact, cell-extracellular matrix interaction and signalling via soluble factors are presented. The advantages of stem cell co-culture strategies and their applications in tissue engineering and regenerative medicine are portrayed through specific examples for several tissues, including orthopaedic soft tissues, bone, heart, vasculature, lung, kidney, liver and nerve. A concise review of the progress and the lessons learned are provided, with a focus on recent developments and their implications. It is hoped that knowledge developed from one tissue can be translated to other tissues. Finally, we address challenges in tissue engineering and regenerative medicine that can potentially be overcome via employing strategies for stem cell co-culture use. Copyright © 2014 John Wiley & Sons, Ltd.
-
Stimulation of DNA synthesis in cultured rat alveolar type II cells
International Nuclear Information System (INIS)
Leslie, C.C.; McCormick-Shannon, K.; Robinson, P.C.; Mason, R.J.
1985-01-01
Restoration of the alveolar epithelium after injury is thought to be dependent on the proliferation of alveolar type II cells. To understand the factors that may be involved in promoting type II cell proliferation in vivo, we determined the effect of potential mitogens and culture substrata on DNA synthesis in rat alveolar type II cells in primary culture. Type II cells cultured in basal medium containing 10% fetal bovine serum (FBS) exhibited essentially no DNA synthesis. Factors that stimulated 3 H-thymidine incorporation included cholera toxin, epidermal growth factor, and rat serum. The greatest degree of stimulation was achieved by plating type II cells on an extracellular matrix prepared from bovine corneal endothelial cells and then by culturing the pneumocytes in medium containing rat serum, cholera toxin, insulin, and epidermal growth factor. Under conditions of stimulation of 3 H-thymidine incorporation there was an increased DNA content per culture dish but no increase in cell number. The ability of various culture conditions to promote DNA synthesis in type II cells was verified by autoradiography. Type II cells were identified by the presence of cytoplasmic inclusions, which were visualized by tannic acid staining before autoradiography. These results demonstrate the importance of soluble factors and culture substratum in stimulating DNA synthesis in rat alveolar type II cells in primary culture
-
Visual cultures in science and technology a comparative history
Hentschel, Klaus
2014-01-01
This book attempts a synthesis. It delves into the rich reservoir of case studies on visual representations in scientific and technological practice that have been accumulated over the past couple of decades by historians, sociologists, and philosophers of science. The main aim is thus located on the meta-level. It adopts an integrative view of recurrently noted general features of visual cultures in science and technology, something hitherto unachieved and believed by many to be a mission impossible. By systematic comparison of numerous case studies, the purview broadens away from myopic microanalysis in search of overriding patterns. The many different disciplines and research areas involved encompass mathematics, technology, natural history, medicine, the geosciences, astronomy, chemistry, and physics. The chosen examples span the period from the Renaissance to the late 20th century. Some pioneers of new visual cultures are portrayed, along with the modes of skill transfer and development. The broad range ...
-
Pursuing Darwin’s curious parallel: Prospects for a science of cultural evolution
2017-01-01
In the past few decades, scholars from several disciplines have pursued the curious parallel noted by Darwin between the genetic evolution of species and the cultural evolution of beliefs, skills, knowledge, languages, institutions, and other forms of socially transmitted information. Here, I review current progress in the pursuit of an evolutionary science of culture that is grounded in both biological and evolutionary theory, but also treats culture as more than a proximate mechanism that is directly controlled by genes. Both genetic and cultural evolution can be described as systems of inherited variation that change over time in response to processes such as selection, migration, and drift. Appropriate differences between genetic and cultural change are taken seriously, such as the possibility in the latter of nonrandomly guided variation or transformation, blending inheritance, and one-to-many transmission. The foundation of cultural evolution was laid in the late 20th century with population-genetic style models of cultural microevolution, and the use of phylogenetic methods to reconstruct cultural macroevolution. Since then, there have been major efforts to understand the sociocognitive mechanisms underlying cumulative cultural evolution, the consequences of demography on cultural evolution, the empirical validity of assumed social learning biases, the relative role of transformative and selective processes, and the use of quantitative phylogenetic and multilevel selection models to understand past and present dynamics of society-level change. I conclude by highlighting the interdisciplinary challenges of studying cultural evolution, including its relation to the traditional social sciences and humanities. PMID:28739929
-
EXPLANTATION OF MESANGIAL CELL HILLOCKS - A METHOD FOR OBTAINING HUMAN MESANGIAL CELLS IN CULTURE
MULLER, EW; KIM, Y; MICHAEL, AF; VERNIER, RL; VANDERHEM, GK; VANDERWOUDE, FJ
A simple method is presented for selective cell culture of human mesangial cells using explanatation of mesangial cell hillocks. Glomeruli which had been incubated with collagenase were explanted on plastic tissue culture flasks. Three to 6 weeks after explantation, a rapidly growing multilayer of
-
International Nuclear Information System (INIS)
Fujitake, Hideki; Okamoto, Yuruko; Okubo, Hiroshi; Miyanomae, Takeshi; Kumagai, Keiko; Mori, K.J.
1981-01-01
Effects of cell concentrations on the survival and repopulation of haemopoietic stem cells after irradiation were studied in the long-term culture of mouse bone marrow cells in vitro. No difference was observed in the survival of the stem cells among cultures in which 0 - 10 7 cells were re-inoculated on the adherent cell colonies in the culture flask. Stem cells showed a significant proliferation within 1 week and the number of the stem cells exceeded the control in 3 weeks after irradiation in the cultures with less than 10 6 re-inoculated cells per flask. In contrast, there was a considerable delay in the onset of stem cell proliferation after irradiation in the culture with 10 7 cells per flask. Based on these results, a possibility that a stimulator of stem cell proliferation, released from irradiated stromal cells, is cancelled by an inhibitory factor produced by irradiated or unirradiated haemopoietic cells is postulated. (author)
-
Vu Thu Hang, N.
2014-01-01
This study is about the application of social constructivism in primary science curriculum in Confucian heritage culture. It was found that the implementation of social constructivism in Confucian heritage culture was low and influenced by cultural divergences between Confucian cultural philosophy
-
2010-04-01
... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Tissue culture media for human ex vivo tissue and cell culture processing applications. 876.5885 Section 876.5885 Food and Drugs FOOD AND DRUG... DEVICES Therapeutic Devices § 876.5885 Tissue culture media for human ex vivo tissue and cell culture...
-
Retinal pigment epithelium culture;a potential source of retinal stem cells.
Akrami, Hassan; Soheili, Zahra-Soheila; Khalooghi, Keynoush; Ahmadieh, Hamid; Rezaie-Kanavi, Mojgan; Samiei, Shahram; Davari, Malihe; Ghaderi, Shima; Sanie-Jahromi, Fatemeh
2009-07-01
To establish human retinal pigment epithelial (RPE) cell culture as a source for cell replacement therapy in ocular diseases. Human cadaver globes were used to isolate RPE cells. Each globe was cut into several pieces of a few millimeters in size. After removing the sclera and choroid, remaining tissues were washed in phosphate buffer saline and RPE cells were isolated using dispase enzyme solution and cultured in Dulbecco's Modified Eagle's Medium: Nutrient Mixture F-12 supplemented with 10% fetal calf serum. Primary cultures of RPE cells were established and spheroid colonies related to progenitor/stem cells developed in a number of cultures. The colonies included purely pigmented or mixed pigmented and non-pigmented cells. After multiple cellular passages, several types of photoreceptors and neural-like cells were detected morphologically. Cellular plasticity in RPE cell cultures revealed promising results in terms of generation of stem/progenitor cells from human RPE cells. Whether the spheroids and neural-like retinal cells were directly derived from retinal stem cells or offspring of trans-differentiating or de-differentiating RPE cells remains to be answered.
-
Enhanced infectivity of bluetongue virus in cell culture by centrifugation.
Sundin, D R; Mecham, J O
1989-01-01
The effects of centrifugation of the infection of cell culture with bluetongue virus (BTV) were investigated. Baby hamster kidney cells were infected with BTV with or without centrifugation. Viral antigen was detected by immunofluorescence at 24 h in both centrifuged and noncentrifuged cultures. However, after 24 h of infection, the production of PFU in centrifuged cell cultures was 10- to 20-fold greater than that seen in cultures not centrifuged. In addition, centrifugation enhanced the dir...
-
Bayati, Vahid; Gazor, Rohoullah; Nejatbakhsh, Reza; Negad Dehbashi, Fereshteh
2016-01-01
As stem cells play a critical role in tissue repair, their manipulation for being applied in regenerative medicine is of great importance. Skin-derived precursors (SKPs) may be good candidates for use in cell-based therapy as the only neural stem cells which can be isolated from an accessible tissue, skin. Herein, we presented a simple protocol to enrich neural SKPs by monolayer adherent cultivation to prove the efficacy of this method. To enrich neural SKPs from dermal cell populations, we have found that a monolayer adherent cultivation helps to increase the numbers of neural precursor cells. Indeed, we have cultured dermal cells as monolayer under serum-supplemented (control) and serum-supplemented culture, followed by serum free cultivation (test) and compared. Finally, protein markers of SKPs were assessed and compared in both experimental groups and differentiation potential was evaluated in enriched culture. The cells of enriched culture concurrently expressed fibronectin, vimentin and nestin, an intermediate filament protein expressed in neural and skeletal muscle precursors as compared to control culture. In addition, they possessed a multipotential capacity to differentiate into neurogenic, glial, adipogenic, osteogenic and skeletal myogenic cell lineages. It was concluded that serum-free adherent culture reinforced by growth factors have been shown to be effective on proliferation of skin-derived neural precursor cells (skin-NPCs) and drive their selective and rapid expansion.
-
Bradley, Jillian H; Stein, Rachel; Randolph, Brad; Molina, Emily; Arnold, Jennifer P; Gregg, Randal K
2017-11-01
Immune impairment mediated by microgravity threatens the success of space exploration requiring long-duration spaceflight. The cells of most concern, T lymphocytes, coordinate the host response against microbial and cancerous challenges leading to elimination and long-term protection. T cells are activated upon recognition of specific microbial peptides bound on the surface of antigen presenting cells, such as dendritic cells (DC). Subsequently, this engagement results in T cell proliferation and differentiation into effector T cells driven by autocrine interleukin-2 (IL-2) and other cytokines. Finally, the effector T cells acquire the weaponry needed to destroy microbial invaders and tumors. Studies conducted on T cells during spaceflight, or using Earth-based culture systems, have shown reduced production of cytokines, proliferation and effector functions as compared to controls. This may account for the cases of viral reactivation events and opportunistic infections associated with astronauts of numerous missions. This work has largely been based upon the outcome of T cell activation by stimulatory factors that target select T cell signaling pathways rather than the complex, signaling events related to the natural process of antigen presentation by DC. This study tested the response of an ovalbumin peptide-specific T cell line, OT-II TCH, to activation by DC when the T cells were cultured 24-120 h in a simulated microgravity (SMG) environment generated by a rotary cell culture system. Following 72 h culture of T cells in SMG (SMG-T) or control static (Static-T) conditions, IL-2 production by the T cells was reduced in SMG-T cells compared to Static-T cells upon stimulation by phorbol 12-myristate 13-acetate (PMA) and ionomycin. However, when the SMG-T cells were stimulated with DC and peptide, IL-2 was significantly increased compared to Static-T cells. Such enhanced IL-2 production by SMG-T cells peaked at 72 h SMG culture time and decreased thereafter
-
Sabanović, Selma
2014-06-01
Using interviews, participant observation, and published documents, this article analyzes the co-construction of robotics and culture in Japan through the technical discourse and practices of robotics researchers. Three cases from current robotics research--the seal-like robot PARO, the Humanoid Robotics Project HRP-2 humanoid, and 'kansei robotics' - show the different ways in which scientists invoke culture to provide epistemological grounding and possibilities for social acceptance of their work. These examples show how the production and consumption of social robotic technologies are associated with traditional crafts and values, how roboticists negotiate among social, technical, and cultural constraints while designing robots, and how humans and robots are constructed as cultural subjects in social robotics discourse. The conceptual focus is on the repeated assembly of cultural models of social behavior, organization, cognition, and technology through roboticists' narratives about the development of advanced robotic technologies. This article provides a picture of robotics as the dynamic construction of technology and culture and concludes with a discussion of the limits and possibilities of this vision in promoting a culturally situated understanding of technology and a multicultural view of science.
-
Khine, Myint Swe; Fisher, Darrell L.
2004-01-01
The purpose of this study was to examine interpersonal behaviour in psychosocial learning environments and to determine the associations between science students' perceptions of their interactions with their teachers, the cultural background of teachers and their attitudinal outcomes. A sample of 1188 students completed the Questionnaire on Teacher Interaction instrument. The responses to two subscales of Test of Science-related Attitudes were used as attitudinal measures. Significant associations between students' perceptions of teacher interpersonal behaviour and the cultural background of teachers were detected. The results showed that students perceived a more favourable interpersonal relationship with Western teachers in the secondary science classrooms. The students in the classes of Western teachers indicated that they enjoyed science lessons more than those in the classes of Asian teachers. Some implications for science instruction in this context are discussed.
-
International Nuclear Information System (INIS)
Nobuhisa, Ikuo; Ohtsu, Naoki; Okada, Seiji; Nakagata, Naomi; Taga, Tetsuya
2007-01-01
The aorta-gonad-mesonephros (AGM) region is a primary source of definitive hematopoietic cells in the midgestation mouse embryo. In cultures of dispersed AGM regions, adherent cells containing endothelial cells are observed first, and then non-adherent hematopoietic cells are produced. Here we report on the characterization of hematopoietic cells that emerge in the AGM culture. Based on the expression profiles of CD45 and c-Kit, we defined three cell populations: CD45 low c-Kit + cells that had the ability to form hematopoietic cell colonies in methylcellulose media and in co-cultures with stromal cells; CD45 low c-Kit - cells that showed a granulocyte morphology; CD45 high c-Kit low/- that exhibited a macrophage morphology. In co-cultures of OP9 stromal cells and freshly prepared AGM cultures, CD45 low c-Kit + cells from the AGM culture had the abilities to reproduce CD45 low c-Kit + cells and differentiate into CD45 low c-Kit - and CD45 high c-Kit low/- cells, whereas CD45 low c-Kit - and CD45 high c-Kit low/- did not produce CD45 low c-Kit + cells. Furthermore, CD45 low c-Kit + cells displayed a long-term repopulating activity in adult hematopoietic tissue when transplanted into the liver of irradiated newborn mice. These results indicate that CD45 low c-Kit + cells from the AGM culture have the potential to reconstitute multi-lineage hematopoietic cells
-
DEFF Research Database (Denmark)
Høyen, Marianne; Mumiah, Rasmusen
the humanities and natural sciences influence the newly educated teachers’ understanding of the teaching profession. From earlier research on teachers in natural science subjects it became clear that teachers from the two major areas are in conflict. Mutual understanding is lacking; the organization...... of the consequences was that teacher students today must choose between to teach either language and literature or maths and therefore, and as a consequence, early in their studies choose between the main areas of culture and nature. Starting from this basis, we want to see if, and in which ways, perspectives from...... of the school day gives priority to cultural subjects; the physical design of the school implies that natural science subjects are of a special kind. and consequently teachers within cultural subjects appear to regard natural science subjects as peripheral educationally to pupils development. Our starting point...
-
Trapped between the two cultures: Urban college students' attitudes toward science
Dawson, Roy Edward
Most Americans agree that science plays an important part in maintaining our leadership role in economics, health, and security. Yet when it comes to science and math we appear to be baffled. Only 25% of Americans understand the process of science well enough to make informed judgment about scientific research reported in the media (National Science Foundation, 1998). What is it that turns Americans away from science? Is it our culture, schools, families, or friends? This study investigates urban college students' attitudes toward science to determine what changes might promote increased participation in the questions, ethical implications and culture of science. Volunteers completed a science questionnaire which included multiple-choice and open-answer questions. The questions were divided into the categories of individual characteristics, home/family, peers, and school/teachers. The multiple-choice questions were analyzed with quantitative statistical techniques. The open-answer questions were used to rate each student's attitude toward science and then analyzed with qualitative methods. Thirteen factors were significant in predicting science attitude but none of them, by itself, explained a large amount of variation. A multiple regression model indicated that the significant factors (in order of importance) were watching science television with your family, having a father not employed in science, having friends who like science, and imagining yourself to be a successful student. A hierarchical multiple regression analysis indicated that the categories of individual characteristics, family, and peers were all significant contributors to the model's prediction of science attitude. School environment/teachers did not add significant predictive power to the model. The qualitative results indicated that the factors of (1) a student's previous experience in science classes and (2) the curriculum philosophy which his or her science teachers employed appeared to be the
-
Ikeda, Kazuhiro; Nagata, Shogo; Okitsu, Teru; Takeuchi, Shoji
2017-06-06
Human pluripotent stem cells are a potentially powerful cellular resource for application in regenerative medicine. Because such applications require large numbers of human pluripotent stem cell-derived cells, a scalable culture system of human pluripotent stem cell needs to be developed. Several suspension culture systems for human pluripotent stem cell expansion exist; however, it is difficult to control the thickness of cell aggregations in these systems, leading to increased cell death likely caused by limited diffusion of gases and nutrients into the aggregations. Here, we describe a scalable culture system using the cell fiber technology for the expansion of human induced pluripotent stem (iPS) cells. The cells were encapsulated and cultured within the core region of core-shell hydrogel microfibers, resulting in the formation of rod-shaped or fiber-shaped cell aggregations with sustained thickness and high viability. By encapsulating the cells with type I collagen, we demonstrated a long-term culture of the cells by serial passaging at a high expansion rate (14-fold in four days) while retaining its pluripotency. Therefore, our culture system could be used for large-scale expansion of human pluripotent stem cells for use in regenerative medicine.
-
Cystine uptake by cultured cells originating from dog proximal tubule segments
International Nuclear Information System (INIS)
States, B.; Reynolds, R.; Lee, J.; Segal, S.
1990-01-01
Large numbers of kidney epithelial cells were cultured successfully from isolated dog proximal tubule segments. Cells in primary culture and in first passage retained the cystine-dibasic amino acid co-transporter system which is found in vivo and in freshly isolated proximal tubule segments. In contrast to other cultured cells, the cystine-glutamate anti-porter was absent in primary cultures. However, this anti-porter system seemed to be developing in cells in first passage. The intracellular ratio of cysteine:reduced glutathione (CSH:GSH) was maintained at 1:36 in both primary cultures and in low passage cells. Incubation of cells in primary culture for 5 min at 37 degrees C with 0.025 mM [ 35 S]L-cystine resulted in incorporation of approximately 36 and 8.5% of the label into intracellular CSH and GSH, respectively. These cultured cells, therefore, seem to be an excellent model system for the eventual elucidation of (a) the inticacies of cystine metabolism and (b) regulation of (1) the cystine-dibasic amino acid co-transporter system and (2) the development of the cysteine-glutamate anti-porter system
-
Learning of science concepts within a traditional socio-cultural ...
African Journals Online (AJOL)
The learning of science concepts within a traditional socio-cultural environment were investigated by looking at: 1) the nature of \\"cognitive border crossing\\" exhibited by the students from the traditional to the scientific worldview, and 2) whether or not three learning theories / hypotheses: border crossing, collaterality, and ...
-
Inhibition of apoptosis using exosomes in Chinese hamster ovary cell culture.
Han, Seora; Rhee, Won Jong
2018-05-01
Animal cell culture technology for therapeutic protein production has shown significant improvement over the last few decades. Chinese hamster ovary (CHO) cells have been widely adapted for the production of biopharmaceutical drugs. In the biopharmaceutical industry, it is crucial to develop cell culture media and culturing conditions to achieve the highest productivity and quality. However, CHO cells are significantly affected by apoptosis in the bioreactors, resulting in a substantial decrease in product quantity and quality. Thus, to overcome the obstacle of apoptosis in CHO cell culture, it is critical to develop a novel method that does not have minimal concern of safety or cost. Herein, we showed for the first time that exosomes, which are nano-sized extracellular vesicles, derived from CHO cells inhibited apoptosis in CHO cell culture when supplemented to the culture medium. Flow cytometric and microscopic analyses revealed that substantial amounts of exosomes were delivered to CHO cells. Higher cell viability after staurosporine treatment was observed by exosome supplementation (67.3%) as compared to control (41.1%). Furthermore, exosomes prevented the mitochondrial membrane potential loss and caspase-3 activation, meaning that the exosomes enhanced cellular activities under pro-apoptotic condition. As the exosomes supplements are derived from CHO cells themselves, it is not only beneficial for the biopharmaceutical productivity of CHO cell culture to inhibit apoptosis, but also from a regulatory standpoint to diminish any safety concerns. Thus, we conclude that the method developed in this research may contribute to the biopharmaceutical industry where minimizing apoptosis in CHO cell culture is beneficial. © 2018 Wiley Periodicals, Inc.
-
Isolation and Characterization of Poliovirus in Cell Culture Systems.
Thorley, Bruce R; Roberts, Jason A
2016-01-01
The isolation and characterization of enteroviruses by cell culture was accepted as the "gold standard" by clinical virology laboratories. Methods for the direct detection of all enteroviruses by reverse transcription polymerase chain reaction, targeting a conserved region of the genome, have largely supplanted cell culture as the principal diagnostic procedure. However, the World Health Organization's Global Polio Eradication Initiative continues to rely upon cell culture to isolate poliovirus due to the lack of a reliable sensitive genetic test for direct typing of enteroviruses from clinical specimens. Poliovirus is able to infect a wide range of mammalian cell lines, with CD155 identified as the primary human receptor for all three seroytpes, and virus replication leads to an observable cytopathic effect. Inoculation of cell lines with extracts of clinical specimens and subsequent passaging of the cells leads to an increased virus titre. Cultured isolates of poliovirus are suitable for testing by a variety of methods and remain viable for years when stored at low temperature.This chapter describes general procedures for establishing a cell bank and routine passaging of cell lines. While the sections on specimen preparation and virus isolation focus on poliovirus, the protocols are suitable for other enteroviruses.
-
A proposal for measurement of science and innovation culture
Energy Technology Data Exchange (ETDEWEB)
Okamura, A
2016-07-01
Why do perceptions about the negative and positive aspects of science, technology, and innovation differ among individuals and across countries? What types of technology do we fear and what types do we embrace? Amongst the general population, which group is most comfortable with new technology and which group is most sceptical about its diffusion? Why are scientific careers popular in some countries and not in others? In the end, is there any relationship between appreciation for science and well-being? How is our relationship with technology linked to national competence and national innovation systems? These questions are of particular importance for science, technology, and innovation policy these days, as shown in some increasingly used policy concepts and keywords, such as ‘responsible research and innovation’, ‘societal impact of science’, ‘science and society’, and ‘innovation for societal issues’. As science and innovation activities are globalized, these ‘cultural’ factors have also gained global importance. In light of the importance of science and innovation culture as a foundation of science, technology and innovation policymaking, a future research aggenda to advance our understanding and measurement is proposed. (Author)
-
SAHA-induced TRAIL-sensitisation of Multiple Myeloma cells is enhanced in 3D cell culture.
Arhoma, A; Chantry, A D; Haywood-Small, S L; Cross, N A
2017-11-15
Multiple Myeloma (MM) is currently incurable despite many novel therapies. Tumour Necrosis Factor-Related Apoptosis-Inducing Ligand (TRAIL) is a potential anti-tumour agent although effects as a single agent are limited. In this study, we investigated whether the Histone Deacetylase (HDAC) inhibitor SAHA can enhance TRAIL-induced apoptosis and target TRAIL resistance in both suspension culture, and 3D cell culture as a model of disseminated MM lesions that form in bone. The effects of SAHA and/or TRAIL in 6 Multiple Myeloma cell lines were assessed in both suspension cultures and in an Alginate-based 3D cell culture model. The effect of SAHA and/or TRAIL was assessed on apoptosis by assessment of nuclear morphology using Hoechst 33342/Propidium Iodide staining. Viable cell number was assessed by CellTiter-Glo luminescence assay, Caspase-8 and -9 activities were measured by Caspase-Glo™ assay kit. TRAIL-resistant cells were generated by culture of RPMI 8226 and NCI-H929 by acute exposure to TRAIL followed by selection of TRAIL-resistant cells. TRAIL significantly induced apoptosis in a dose-dependent manner in OPM-2, RPMI 8226, NCI-H929, U266, JJN-3 MM cell lines and ADC-1 plasma cell leukaemia cells. SAHA amplified TRAIL responses in all lines except OPM-2, and enhanced TRAIL responses were both via Caspase-8 and -9. SAHA treatment induced growth inhibition that further increased in the combination treatment with TRAIL in MM cells. The co-treatment of TRAIL and SAHA reduced viable cell numbers all cell lines. TRAIL responses were further potentiated by SAHA in 3D cell culture in NCI-H929, RPMI 8226 and U266 at lower TRAIL + SAHA doses than in suspension culture. However TRAIL responses in cells that had been selected for TRAIL resistance were not further enhanced by SAHA treatment. SAHA is a potent sensitizer of TRAIL responses in both TRAIL sensitive and resistant cell lines, in both suspension and 3D culture, however SAHA did not sensitise TRAIL-sensitive cell
-
Jackson, Matthew C.; Galvez, Gino; Landa, Isidro; Buonora, Paul; Thoman, Dustin B.
2016-01-01
Recent research suggests that underrepresented minority (URM) college students, and especially first-generation URMs, may lose motivation to persist if they see science careers as unable to fulfill culturally relevant career goals. In the present study, we used a mixed-methods approach to explore patterns of motivation to pursue physical and life…
-
Flux analysis of mammalian cell culture
Martens, D.E.; Tramper, J.
2010-01-01
Animal cells are used for the production of vaccines and pharmaceutical proteins. The increase in demand for these products requires an increase in volumetric productivity of animal cell culture processes, which can be attained through an increase in biomass concentration and/or specific
-
Reforming science: methodological and cultural reforms.
Casadevall, Arturo; Fang, Ferric C
2012-03-01
Contemporary science has brought about technological advances and an unprecedented understanding of the natural world. However, there are signs of dysfunction in the scientific community as well as threats from diverse antiscience and political forces. Incentives in the current system place scientists under tremendous stress, discourage cooperation, encourage poor scientific practices, and deter new talent from entering the field. It is time for a discussion of how the scientific enterprise can be reformed to become more effective and robust. Serious reform will require more consistent methodological rigor and a transformation of the current hypercompetitive scientific culture.
-
Hall, R.D.
2000-01-01
The use of cultured plant cells in either organized or unorganized form has increased vey considerably in the last 10-15 yr. Many new technologies have been developed and applications in both fundamental and applied research have led to the development of some powerful tools for improving our
-
Directory of Open Access Journals (Sweden)
Ruth Olmer
2018-05-01
Full Text Available Summary: Endothelial cells (ECs are involved in a variety of cellular responses. As multifunctional components of vascular structures, endothelial (progenitor cells have been utilized in cellular therapies and are required as an important cellular component of engineered tissue constructs and in vitro disease models. Although primary ECs from different sources are readily isolated and expanded, cell quantity and quality in terms of functionality and karyotype stability is limited. ECs derived from human induced pluripotent stem cells (hiPSCs represent an alternative and potentially superior cell source, but traditional culture approaches and 2D differentiation protocols hardly allow for production of large cell numbers. Aiming at the production of ECs, we have developed a robust approach for efficient endothelial differentiation of hiPSCs in scalable suspension culture. The established protocol results in relevant numbers of ECs for regenerative approaches and industrial applications that show in vitro proliferation capacity and a high degree of chromosomal stability. : In this article, U. Martin and colleagues show the generation of hiPSC endothelial cells in scalable cultures in up to 100 mL culture volume. The generated ECs show in vitro proliferation capacity and a high degree of chromosomal stability after in vitro expansion. The established protocol allows to generate hiPSC-derived ECs in relevant numbers for regenerative approaches. Keywords: hiPSC differentiation, endothelial cells, scalable culture
-
Electrospinning PCL Scaffolds Manufacture for Three-Dimensional Breast Cancer Cell Culture
Directory of Open Access Journals (Sweden)
Marc Rabionet
2017-08-01
Full Text Available In vitro cell culture is traditionally performed within two-dimensional (2D environments, providing a quick and cheap way to study cell properties in a laboratory. However, 2D systems differ from the in vivo environment and may not mimic the physiological cell behavior realistically. For instance, 2D culture models are thought to induce cancer stem cells (CSCs differentiation, a rare cancer cell subpopulation responsible for tumor initiation and relapse. This fact hinders the development of therapeutic strategies for tumors with a high relapse percentage, such as triple negative breast cancer (TNBC. Thus, three-dimensional (3D scaffolds have emerged as an attractive alternative to monolayer culture, simulating the extracellular matrix structure and maintaining the differentiation state of cells. In this work, scaffolds were fabricated through electrospinning different poly(ε-caprolactone-acetone solutions. Poly(ε-caprolactone (PCL meshes were seeded with triple negative breast cancer (TNBC cells and 15% PCL scaffolds displayed significantly (p < 0.05 higher cell proliferation and elongation than the other culture systems. Moreover, cells cultured on PCL scaffolds exhibited higher mammosphere forming capacity and aldehyde dehydrogenase activity than 2D-cultured cells, indicating a breast CSCs enrichment. These results prove the powerful capability of electrospinning technology in terms of poly(ε-caprolactone nanofibers fabrication. In addition, this study has demonstrated that electrospun 15% PCL scaffolds are suitable tools to culture breast cancer cells in a more physiological way and to expand the niche of breast CSCs. In conclusion, three-dimensional cell culture using PCL scaffolds could be useful to study cancer stem cell behavior and may also trigger the development of new specific targets against such malignant subpopulation.
-
Directory of Open Access Journals (Sweden)
Mee-Hae Kim
2017-10-01
Full Text Available In cell manufacturing, the establishment of a fully automated, microfluidic, cell culture system that can be used for long-term cell cultures, as well as for process optimization is highly desirable. This study reports the development of a novel chip bioreactor system that can be used for automated long-term maturation cultures of retinal pigment epithelial (RPE cells. The system consists of an incubation unit, a medium supply unit, a culture observation unit, and a control unit. In the incubation unit, the chip contains a closed culture vessel (2.5 mm diameter, working volume 9.1 μL, which can be set to 37 °C and 5% CO2, and uses a gas-permeable resin (poly- dimethylsiloxane as the vessel wall. RPE cells were seeded at 5.0 × 104 cells/cm2 and the medium was changed every day by introducing fresh medium using the medium supply unit. Culture solutions were stored either in the refrigerator or the freezer, and fresh medium was prepared before any medium change by warming to 37 °C and mixing. Automated culture was allowed to continue for 30 days to allow maturation of the RPE cells. This chip culture system allows for the long-term, bubble-free, culture of RPE cells, while also being able to observe cells in order to elucidate their cell morphology or show the presence of tight junctions. This culture system, along with an integrated on-line monitoring system, can therefore be applied to long-term cultures of RPE cells, and should contribute to process control in RPE cell manufacturing.
-
Wei, Wei; Xu, Chao; Ye, Zhi-Yong; Huang, Xiao-Jun; Yuan, Jia-En; Ma, Tian-Bao; Lin, Han-Biao; Chen, Xiu-Qiong
2016-10-25
The aim of the present study was to obtain the qualified hematopoietic stem/progenitor cells (HSC/HPC) and human umbilical cord-mesenchymal stem cells (MSC) in vitro in the co-culture system. Cord blood mononuclear cells were separated from umbilical cord blood by Ficoll lymphocyte separation medium, and then CD34 + HSC was collected by MACS immunomagnetic beads. The selected CD34 + HSC/HPC and MSC were transferred into culture flask. IMDM culture medium with 15% AB-type cord plasma supplemented with interleukin-3 (IL-3), IL-6, thrombopoietin (TPO), stem cell factor (SCF) and FMS-like tyrosine kinase 3 ligand (Flt-3L) factors were used as the co-culture system for the amplification of HSC/HPC and MSC. The cellular growth status and proliferation on day 6 and 10 after co-culture were observed by using inverted microscope. The percentage of positive expression of CD34 in HSC/HPC, as well as the percentages of positive expressions of CD105, CD90, CD73, CD45, CD34 and HLA-DR in the 4 th generation MSC, was tested by flow cytometry. Semisolid colony culture was used to test the HSC/HPC colony forming ability. The osteogenic, chondrogenesis and adipogenic ability of the 4 th generation MSC were assessed. The karyotype analysis of MSC was conducted by colchicines. The results demonstrated that the HSC/HPC of co-culture group showed higher ability of amplification, CFU-GM and higher CD34 + percentage compared with the control group. The co-cultured MSC maintained the ability to differentiate into bone cells, fat cells and chondrocytes. And the karyotype stability of MSC remained normal. These results reveal that the appropriate co-culture system for MSC and HSC is developed, and via this co-culture system we could gain both two kinds of these cells. The MSCs under the co-culture system maintain the biological characteristics. The CFU-GM ability, cell counting and the flow cytometry results of HSC/HPC under the co-culture system are conform to the criterion, showing that
-
The Teleodynamics of Language, Culture, Technology and Science (LCT&S
Directory of Open Access Journals (Sweden)
Robert K. Logan
2013-02-01
Full Text Available Logan [1] in his book The Extended Mind developed the hypothesis that language, culture, technology and science can be treated as organisms that evolve and reproduce themselves. This idea is extended by making use of the notion of teleodynamics that Deacon [2] introduced and developed in his book Incomplete Nature to explain the nature of life, sentience, mind and a self that acts in its own interest. It is suggested that language, culture, technology and science (LCT&S like living organisms also act in their own self-interest, are self-correcting and are to a certain degree autonomous even though they are obligate symbionts with their human hosts. Specifically, it will be argued that LCT&S are essentially teleodynamic systems, which Deacon defines as “self-creating, self-maintaining, self-reproducing, individuated systems [2] (p. 325”.
-
2013-08-16
....R50000] Notice of Intent To Repatriate Cultural Item: Rochester Museum & Science Center, Rochester, NY AGENCY: National Park Service, Interior. ACTION: Notice. SUMMARY: The Rochester Museum & Science Center... that the cultural item listed in this notice meets the definition of a sacred object and an object of...
-
Energy Technology Data Exchange (ETDEWEB)
Bulard, C; Mary, J; Chaumont, D; Gudin, C
1982-11-01
Myrtillocactus tissue cultures are grown from the epicotyl of young plantlets. With an appropriate growing medium it is possible, after transfer of fragments of these cultures to a liquid environment, to obtain dissociation and proliferation of cells. The production of betalaic pigments is induced in solid surroundings by adjustement of the growing medium composition and can be maintained in a liquid environment. The multiplication of pigmented cells in suspension may thus be obtained. The conversion of Asparagus cell suspensions from the heterotrophic state (use of lactose as source of carbon) to the autotrophic state (carbon supplied by CO/sub 2/) is obtained by a gradual reduction in the sugar concentration of the medium combined with a rise in the CO/sub 2/ content of the gas mixture atmosphere injected into the cultivator. The passage to the autotrophic state of a Myrtillocactus suspension would enable the production conditions of a metabolite (Betalaine) to be studied by micro-algae culture techniques.
-
Hypoxic contraction of cultured pulmonary vascular smooth muscle cells
International Nuclear Information System (INIS)
Murray, T.R.; Chen, L.; Marshall, B.E.; Macarak, E.J.
1990-01-01
The cellular events involved in generating the hypoxic pulmonary vasoconstriction response are not clearly understood, in part because of the multitude of factors that alter pulmonary vascular tone. The goal of the present studies was to determine if a cell culture preparation containing vascular smooth muscle (VSM) cells could be made to contract when exposed to a hypoxic atmosphere. Cultures containing only fetal bovine pulmonary artery VSM cells were assessed for contractile responses to hypoxic stimuli by two methods. In the first, tension forces generated by cells grown on a flexible growth surface (polymerized polydimethyl siloxane) were manifested as wrinkles and distortions of the surface under the cells. Wrinkling of the surface was noted to progressively increase with time as the culture medium bathing the cells was made hypoxic (PO2 approximately 25 mmHg). The changes were sometimes reversible upon return to normoxic conditions and appeared to be enhanced in cells already exhibiting evidence of some baseline tone. Repeated passage in culture did not diminish the hypoxic response. Evidence for contractile responses to hypoxia was also obtained from measurements of myosin light chain (MLC) phosphorylation. Conversion of MLC to the phosphorylated species is an early step in the activation of smooth muscle contraction. Lowering the PO2 in the culture medium to 59 mmHg caused a 45% increase in the proportion of MLC in the phosphorylated form as determined by two-dimensional gel electrophoresis. Similarly, cultures preincubated for 4 h with 32P and then exposed to normoxia or hypoxia for a 5-min experimental period showed more than twice as much of the label in MLCs of the hypoxic cells
-
Contributions of 3D Cell Cultures for Cancer Research.
Ravi, Maddaly; Ramesh, Aarthi; Pattabhi, Aishwarya
2017-10-01
Cancer cell lines have contributed immensely in understanding the complex physiology of cancers. They are excellent material for studies as they offer homogenous samples without individual variations and can be utilised with ease and flexibility. Also, the number of assays and end-points one can study is almost limitless; with the advantage of improvising, modifying or altering several variables and methods. Literally, a new dimension to cancer research has been achieved by the advent of 3Dimensional (3D) cell culture techniques. This approach increased many folds the ways in which cancer cell lines can be utilised for understanding complex cancer biology. 3D cell culture techniques are now the preferred way of using cancer cell lines to bridge the gap between the 'absolute in vitro' and 'true in vivo'. The aspects of cancer biology that 3D cell culture systems have contributed include morphology, microenvironment, gene and protein expression, invasion/migration/metastasis, angiogenesis, tumour metabolism and drug discovery, testing chemotherapeutic agents, adaptive responses and cancer stem cells. We present here, a comprehensive review on the applications of 3D cell culture systems for these aspects of cancers. J. Cell. Physiol. 232: 2679-2697, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.
-
Evaluation of a dental pulp-derived cell sheet cultured on amniotic membrane substrate.
Honjo, Ken-ichi; Yamamoto, Toshiro; Adachi, Tetsuya; Amemiya, Takeshi; Mazda, Osam; Kanamura, Narisato; Kita, Masakazu
2015-01-01
Mesenchymal stem cells (MSC) are transplanted for periodontal tissue regeneration, and the periodontal ligament (PDL) is regenerated using a cultured cell sheet. This cultured cell sheet is prepared using PDL-derived cells, growth factors, and amniotic membrane (AM). Dental pulp (DP)-derived cells can be easily obtained from extracted wisdom teeth, proliferate rapidly, and are less susceptible to bacterial infection than PDL-derived cells. Thus, to prepare a novel cell sheet, DP-derived cells were cultured on AM as a culture substrate for immunohistochemical examination. Wisdom teeth extracted from three adults were cut along the cement-enamel border. DP tissue was collected, minced, and primarily cultured. After three or four passage cultures, DP-derived cells were cultured on AM, followed by hematoxylin-eosin (H-E) and immunofluorescence staining. DP-derived cells cultured on AM formed a layered structure. Cells positive for vimentin, Ki-67, ZO-1, desmoplakin, CD29, 44, 105 or 146, STRO-1, collagen IV or VII or laminin 5 or α5 chain were localized. DP-derived cells proliferated on AM, while retaining the properties of DP, which allowed the cultured cell sheet to be prepared. In addition, the cultured cell sheet contained MSC, which suggests its potential application in periodontal tissue regeneration.
-
Cytotoxicity of TSP in 3D Agarose Gel Cultured Cell.
Directory of Open Access Journals (Sweden)
Song-I Chun
Full Text Available A reference reagent, 3-(trimethylsilyl propionic-2, 2, 3, 3-d4 acid sodium (TSP, has been used frequently in nuclear magnetic resonance (NMR and magnetic resonance spectroscopy (MRS as an internal reference to identify cell and tissue metabolites, and determine chemical and protein structures. This reference material has been exploited for the quantitative and dynamic analyses of metabolite spectra acquired from cells. The aim of this study was to evaluate the cytotoxicity of TSP on three-dimensionally, agarose gel, cultured cells.A human osteosarcoma cell line (MG-63 was selected, and cells were three dimensionally cultured for two weeks in an agarose gel. The culture system contained a mixture of conventional culture medium and various concentrations (0, 1, 3, 5, 7, 10, 20 30 mM of TSP. A DNA quantification assay was conducted to assess cell proliferation using Quant-iT PicoGreen dsDNA reagent and kit, and cell viability was determined using a LIVE/DEAD Viability/Cytotoxicity kit. Both examinations were performed simultaneously at 1, 3, 7 and 14 days from cell seeding.In this study, the cytotoxicity of TSP in the 3D culture of MG-63 cells was evaluated by quantifying DNA (cell proliferation and cell viability. High concentrations of TSP (from 10 to 30 mM reduced both cell proliferation and viability (to 30% of the control after one week of exposure, but no such effects were found using low concentrations of TSP (0-10 mM.This study shows that low concentrations of TSP in 3D cell culture medium can be used for quantitative NMR or MRS examinations for up to two weeks post exposure.
-
Directory of Open Access Journals (Sweden)
Kim TE
2016-03-01
Full Text Available Tae-Eon Kim,1–3,* Chang Gun Kim,1–3,* Jin Soo Kim,4 Songwan Jin,4 Sik Yoon,5 Hae-Rahn Bae,6 Jeong-Hwa Kim,7,8 Young Hun Jeong,7,8 Jong-Young Kwak1–3 1Department of Pharmacology, School of Medicine, 2Department of Biomedical Sciences, The Graduate School, Ajou University, Suwon, South Korea; 3Immune Network Pioneer Research Center, Ajou University Medical Center, Suwon, South Korea; 4Department of Mechanical Engineering, Korea Polytechnic University, Gyeonggi, South Korea; 5Department of Anatomy, School of Medicine, Pusan National University, Yangsan, South Korea; 6Department of Physiology, College of Medicine, Dong-A University, Busan, South Korea; 7School of Mechanical Engineering, 8Department of Mechanical Engineering, Graduate School, Kyungpook National University, Daegu, South Korea *These authors contributed equally to this work Abstract: An artificial three-dimensional (3D culture system that mimics the tumor microenvironment in vitro is an essential tool for investigating the cross-talk between immune and cancer cells in tumors. In this study, we developed a 3D culture system using an electrospun poly(ε-caprolactone (PCL nanofibrous scaffold (NFS. A hybrid NFS containing an uninterrupted network of nano- and submicron-scale fibers (400 nm to 2 µm was generated by deposition onto a stainless steel mesh instead of an aluminum plate. The hybrid NFS contained multiplanar pores in a 3D structure. Surface-seeded mouse CT26 colon cancer cells and bone marrow-derived dendritic cells (BM-DCs were able to infiltrate the hybrid NFS within several hours. BM-DCs cultured on PCL nanofibers showed a baseline inactive form, and lipopolysaccharide (LPS-activated BM-DCs showed increased expression of CD86 and major histocompatibility complex Class II. Actin and phosphorylated FAK were enriched where unstimulated and LPS-stimulated BM-DCs contacted the fibers in the 3D hybrid NFS. When BM-DCs were cocultured with mitoxantrone-treated CT26 cells in
-
Directory of Open Access Journals (Sweden)
L. Koohi,
2014-07-01
Full Text Available German chamomile (Matricaria chamomilla L. is one of the most important medicinal plants that its essential oils used in different medicinal industries. In this study which was carried out in 2013 growing season at the Faculty of Agricultural Sciences of the University of Mohaghegh Ardabili, the in vitro response of leaf and hypocotyl explants of German Chamomile in B5 medium supplemented with different levels of plant growth regulators including 2,4-D, naphthalene acetic acid (NAA, kinetin and 6-benzylaminopurine (BAP were investigated in a factorial experiment based on completely randomized design (CRD.In addition, cell suspension cultures were established and characterized. Hypocotyl and leaf explants exhibited cell proliferation and produced callus within 1-2 weeks. The highest fresh weight of the callus (264.1 mg was produced by leaf explants in the medium supplemented with 0.5 mg/l 2,4-D and 1 mg/l BAP. However, the leaf explants cultured on medium containing 1.5 mg/l 2,4-D showed the lowest cell proliferation and callus yield (40.42 mg. The highest percentage of root induction from leaf explants (58.73% was observed on the medium containing 4 mg/l 2,4-D and 1 mg/l Kin, and from hypocotyl explants (48.61% was observed on medium supplemented with 1.5 mg/l NAA. The 42.22% of calli derived from hypocotyl explants on B5 medium supplemented with 4 mg/l NAA and 3 mg/l BAP, were friable. Cell suspension cultures of German chamomile were established by transferring of hypocotyl-derived friable calli into the MS medium supplemented with 1.5 mg/l 2,4-D and 1 mg/l kinetin. The growth curve of cell proliferations started 4 days after culture and continued to grow until day 13th, where the cells entered stationary phase.
-
Establishment and characterization of American elm cell suspension cultures
Steven M. Eshita; Joseph C. Kamalay; Vicki M. Gingas; Daniel A. Yaussy
2000-01-01
Cell suspension cultures of Dutch elm disease (DED)-tolerant and DED-susceptible American elms clones have been established and characterized as prerequisites for contrasts of cellular responses to pathogen-derived elicitors. Characteristics of cultured elm cell growth were monitored by A700 and media conductivity. Combined cell growth data for all experiments within a...
-
Bondima, Michelle Harris
This ethnographic in nature study explores how two middle school science teachers who have classes populated by urban African Americans teach their students and how their students perceive their teaching. Since urban African American students continue to perform lower than desired on measures of science achievement, there is an urgent need to understand what pedagogical methodologies assist and hinder urban African American students in achieving higher levels of success in science. A pedagogical methodology that theorists posit assists subordinated school populations is culturally responsive pedagogy. Culturally responsive pedagogy is defined as a teaching methodology concerned with preparing students to question inequality, racism, and injustice. Teachers who use culturally responsive pedagogy respect the culture students bring to the class, and require that the teachers willingly do whatever is necessary to educate students (Nieto, 2000). The teacher participants were two female African Americans who were identified by their school supervisors as being highly effective with urban African American students. The researcher presented the teachers in separate case studies conducted over a data collection period of nine months. Data were collected by participant observation, interviews, and artifact collection. Data were analyzed by application of grounded theory techniques. Findings of the teachers' (and the students') beliefs about pedagogy that both assisted and hindered the students' performance in science were reported in a rich and nuanced storytelling manner based on multiple perspectives (teachers', students', and the researcher's). Pedagogical methodologies that the teachers used that assisted their students were the use of cultural metaphors and images in science and applications of motivational techniques that encouraged a nurturing relationship between the teacher and her students. Pedagogical methodologies that hindered students varied by teacher
-
Directory of Open Access Journals (Sweden)
Imran S. Chaudhry
2011-01-01
Full Text Available The biological effects of only a finite number of tobacco toxins have been studied. Here, we describe exposure of cultures of human bronchial epithelial cells to low concentrations of tobacco carcinogens: nickel sulphate, benzo(bfluoranthene, N-nitrosodiethylamine, and 4-(methylnitrosamino-1-(3-pyridyl-1-butanone (NNK. After a 24-hour exposure, EGFR was expressed in cell membrane and cytoplasm, BCL-2 was expressed only in the irregular nuclei of large atypical cells, MKI67 was expressed in nuclei with no staining in larger cells, cytoplasmic BIRC5 with stronger nuclear staining was seen in large atypical cells, and nuclear TP53 was strongly expressed in all cells. After only a 24-hour exposure, cells exhibited atypical nuclear and cytoplasmic features. After a 48-hour exposure, EGFR staining was localized to the nucleus, BCL-2 was slightly decreased in intensity, BIRC5 was localized to the cytoplasm, and TP53 staining was increased in small and large cells. BCL2L1 was expressed in both the cytoplasm and nuclei of cells at 24- and 48-hour exposures. We illustrate that short-termexposure of a bronchial epithelial cell line to smoking-equivalent concentrations of tobacco carcinogens alters the expression of key proliferation regulatory genes, EGFR, BCL-2, BCL2L1, BIRC5, TP53, and MKI67, similar to that reported in biopsy specimens of pulmonary epithelium described to be preneoplastic lesions.
-
International Nuclear Information System (INIS)
Mori, K.J.; Izumi, Hiroko; Seto, Akira
1981-01-01
A culture system was established in which haemopoietic stem cells can undergo a recovery proliferation after a depletion of the stem cells, completely in vitro. To elucidate the source of the stimulatory factors, normal bone marrow cells were overlayed on top of the irradiated adherent 'stromal' cell colonies in the bone marrow cell culture. This stimulated the proliferation of haemopoietic stem cells in the cultured cells in suspension. The present results indicate that the stromal cells produce factors which stimulate stem cell proliferation. Whether the stimulation is evoked by direct cell-cell interactions or by humoral factors is as yet to be studied. (author)
-
Immunocytochemical characterization of primary cell culture in canine transmissible venereal tumor
Directory of Open Access Journals (Sweden)
Luis M.M. Flórez
Full Text Available Abstract: Immunochemistry with anti-vimentin, anti-lysozyme, anti-alpha 1 antitrypsin, anti-CD3 and anti-CD79α antibodies has been used for characterization of primary cell culture in the transmissible venereal tumor (TVT. Samples for primary cell culture and immunohistochemistry assays were taken from eight dogs with cytological and clinical diagnosis of TVT. To validate the immunochemical results in the primary cell culture of TVT, a chromosome count was performed. For the statistical analysis, the Mann-Whitney test with p<0.05 was used. TVT tissues and culture cells showed intense anti-vimentin immunoreactivity, lightly to moderate immunoreactivity for anti-lysozyme, and mild for anti-alpha-antitrypsin. No marking was achieved for CD3 and CD79α. All culture cells showed chromosomes variable number of 56 to 68. This is the first report on the use of immunocytochemical characterization in cell culture of TVT. Significant statistic difference between immunochemistry in tissue and culture cell was not established, what suggests that the use of this technique may provide greater certainty for the confirmation of tumors in the primary culture. This fact is particularly important because in vitro culture of tumor tissues has been increasingly used to provide quick access to drug efficacy and presents relevant information to identify potential response to anticancer medicine; so it is possible to understand the behavior of the tumor.
-
Irradiated murine fibroblasts as feeder layer used in human cell culture
International Nuclear Information System (INIS)
Almeida, Tiago L.; Klingbeil, Fatima G.; Yoshito, Daniele; Caproni, Priscila; Mathor, Monica B.; Herson, Marisa R.
2007-01-01
In 1975, Rheinwald and Green published an in vitro model for keratinocyte cell cultures in which the use of murine fibroblasts, as a feeder layer was introduced. These cells are modified fibroblasts, which presence render keratinocyte cells to remain proliferative for longer periods of time. This optimization of culture outputs has allowed for several clinical applications of confluent keratinocyte cultures as skin substitutes or wound dressings in situations such as post burn extensive skin loss, loss of oral mucosa, and other skin disorders. Nevertheless, proliferation of fibroblast in co-culture with keratinocytes must be controlled by anti-proliferative measures such as irradiation; at the same time, keratinocytes require specific nutrients in the culture medium, which may interfere with the fibroblast feeder layer viability. Therefore, the thorough understanding of the impact of different issues such as culture media composition, irradiation dose and pre-plating storage conditions of irradiated fibroblast to be used as feeder layer in these co-culture systems is important. In this work, changes as far as viability and proliferative rates of irradiated fibroblasts in culture were evaluated in relation to the type of culture medium used, dose of gamma radiation exposure, storage and timing of cell plating post irradiation. Results indicate that the type of culture medium used and time-lag between irradiation, refrigeration and plating of irradiated cells do not have significant impact in culture outcomes. However, the dose of gamma radiation administered to the cells may influence the final quality of these cells if to be used as a feeder layer. (author)
-
Animal-cell culture media: History, characteristics, and current issues.
Yao, Tatsuma; Asayama, Yuta
2017-04-01
Cell culture technology has spread prolifically within a century, a variety of culture media has been designed. This review goes through the history, characteristics and current issues of animal-cell culture media. A literature search was performed on PubMed and Google Scholar between 1880 and May 2016 using appropriate keywords. At the dawn of cell culture technology, the major components of media were naturally derived products such as serum. The field then gradually shifted to the use of chemical-based synthetic media because naturally derived ingredients have their disadvantages such as large batch-to-batch variation. Today, industrially important cells can be cultured in synthetic media. Nevertheless, the combinations and concentrations of the components in these media remain to be optimized. In addition, serum-containing media are still in general use in the field of basic research. In the fields of assisted reproductive technologies and regenerative medicine, some of the medium components are naturally derived in nearly all instances. Further improvements of culture media are desirable, which will certainly contribute to a reduction in the experimental variation, enhance productivity among biopharmaceuticals, improve treatment outcomes of assisted reproductive technologies, and facilitate implementation and popularization of regenerative medicine.
-
Cell Culture Assay for Human Noroviruses [response
Energy Technology Data Exchange (ETDEWEB)
Straub, Tim M.; Honer Zu Bentrup, Kerstin; Orosz Coghlan, Patricia; Dohnalkova, Alice; Mayer, Brooke K.; Bartholomew, Rachel A.; Valdez, Catherine O.; Bruckner-Lea, Cindy J.; Gerba, Charles P.; Abbaszadegan, Morteza A.; Nickerson, Cheryl A.
2007-07-01
We appreciate the comments provided by Leung et al., in response to our recently published article “In Vitro Cell Culture Infectivity Assay for Human Noroviruses” by Straub et al. (1). The specific aim of our project was to develop an in vitro cell culture infectivity assay for human noroviruses (hNoV) to enhance risk assessments when they are detected in water supplies. Reverse transcription (RT) qualitative or quantitative PCR are the primary assays for waterborne NoV monitoring. However, these assays cannot distinguish between infectious vs. non-infectious virions. When hNoV is detected in water supplies, information provided by our infectivity assay will significantly improve risk assessment models and protect human health, regardless of whether we are propagating NoV. Indeed, in vitro cell culture infectivity assays for the waterborne pathogen Cryptosporidium parvum that supplement approved fluorescent microscopy assays, do not result in amplification of the environmentally resistant hard-walled oocysts (2). However, identification of life cycle stages in cell culture provides evidence of infectious oocysts in a water supply. Nonetheless, Leung et al.’s assertion regarding the suitability of our method for the in vitro propagation of high titers of NoV is valid for the medical research community. In this case, well-characterized challenge pools of virus would be useful for developing and testing diagnostics, therapeutics, and vaccines. As further validation of our published findings, we have now optimized RT quantitative PCR to assess the level of viral production in cell culture, where we are indeed finding significant increases in viral titer. The magnitude and time course of these increases is dependent on both virus strain and multiplicity of infection. We are currently preparing a manuscript that will discuss these findings in greater detail, and the implications this may have for creating viral challenge pools
-
Rheological characteristics of cell suspension and cell culture of Perilla frutescens.
Zhong, J J; Seki, T; Kinoshita, S; Yoshida, T
1992-12-05
Physical properties such as viscosity, fluid dynamic behavior of cell suspension, and size distribution of cell aggregates of a plant, Perilla frustescens, cultured in a liquid medium were studied. As a result of investigations using cells harvester after 12 days of cultivation in a flask, it was found that the apparent viscosity of the cell suspension did not change with any variation of cell concentration below 5 g dry cell/L but markedly increased when the cell concentration increased over 12.8 g dry cell/L. The cell suspension exhibited the characteristics of a Bingham plastic fluid with a small yield stress. The size of cell aggregates in the range 74 to 500 mum did not influence the rheological characteristics of the cell suspension. The rheological characteristics of cultivation mixtures of P. frutescens cultivated in a flask and in a bioreactor were also investigated. The results showed that the flow characteristics of the cell culture could be described by a Bingham plastic model. At the later stage of cultivation, the apparent viscosity increased steadily, even though the biomass concentration (by dry weight) decreased, due to the increase of individual cell size. (c) 1992 John Wiley & Sons, Inc.
-
Science, Linguistique, Littérature : trois disciplines, deux discours, une culture
Directory of Open Access Journals (Sweden)
Sandrine SORLIN
2010-09-01
Full Text Available Cet article propose une réflexion sur les frontières épistémologiques et méthodologiques qui séparent trois disciplines universitaires. Portées par un discours propre à l’économie de leur discipline, la littérature et la science se pensent l’une l’autre comme « deux cultures » antinomiques. Or chaque discipline aborde son objet d’étude à partir d’un paradigme qui est culturellement déterminé. La concomitance de l’apparition des « théories du chaos » en science et du mouvement postmoderne en littérature dans les années 70 par exemple s’explique par leur appartenance à une même culture qui, à un moment donné, a opéré un changement de paradigme informant toutes les disciplines. La linguistique n’y a pas échappé ; pourtant le problème de ses frontières demeure. Nous mettrons en avant la transversalité de cette discipline, laquelle permet - entre autres - de rendre compte non seulement d’un style littéraire mais aussi de ce qu’on peut appeler la « rhétorique » ou la textualité de la science.This paper aims at considering the methodological and epistemological boundaries separating three academic disciplines. Underlain by a discourse that is proper to the economy of their discipline, literature and science regard each other as two opposite cultures. Yet each tackles its object of study through a culturally-determined paradigm. The simultaneous birth of chaos theory in science and postmodern aestheticism in literature in the 70s for instance can only be accounted for if we think of them as belonging to the same culture, which, at some point, brought about a change in paradigm that informed all disciplines. Linguistics underwent the same process. Yet the question of its boundaries remains. We will see to what extent linguistics can be seen as cross-disciplinary, in its study of the rhetoric of both literary and scientific textuality.
-
International Nuclear Information System (INIS)
Zhang, Fenxi; Hong, Yan; Liang, Wenmei; Ren, Tongming; Jing, Suhua; Lin, Juntang
2012-01-01
Highlights: ► Co-culture of Sertoli cells (SCs) with human umbilical cord mesenchymal stem cells (UCMSCs). ► Presence of SCs dramatically increased proliferation and migration of UCMSCs. ► Presence of SCs stimulated expression of Mdm2, Akt, CDC2, Cyclin D, CXCR4, MAPKs. -- Abstract: Human umbilical cord mesenchymal stem cells (hUCMSCs) have been recently used in transplant therapy. The proliferation and migration of MSCs are the determinants of the efficiency of MSC transplant therapy. Sertoli cells are a kind of “nurse” cells that support the development of sperm cells. Recent studies show that Sertoli cells promote proliferation of endothelial cells and neural stem cells in co-culture. We hypothesized that co-culture of UCMSCs with Sertoli cells may also promote proliferation and migration of UCMSCs. To examine this hypothesis, we isolated UCMSCs from human cords and Sertoli cells from mouse testes, and co-cultured them using a Transwell system. We found that UCMSCs exhibited strong proliferation ability and potential to differentiate to other cell lineages such as osteocytes and adipocytes. The presence of Sertoli cells in co-culture significantly enhanced the proliferation and migration potential of UCMSCs (P < 0.01). Moreover, these phenotypic changes were accompanied with upregulation of multiple genes involved in cell proliferation and migration including phospho-Akt, Mdm2, phospho-CDC2, Cyclin D1, Cyclin D3 as well as CXCR4, phospho-p44 MAPK and phospho-p38 MAPK. These findings indicate that Sertoli cells boost UCMSC proliferation and migration potential.
-
Endothelial cells stimulate growth of normal and cancerous breast epithelial cells in 3D culture
Directory of Open Access Journals (Sweden)
Magnusson Magnus K
2010-07-01
Full Text Available Abstract Background Epithelial-stromal interaction provides regulatory signals that maintain correct histoarchitecture and homeostasis in the normal breast and facilitates tumor progression in breast cancer. However, research on the regulatory role of the endothelial component in the normal and malignant breast gland has largely been neglected. The aim of the study was to investigate the effects of endothelial cells on growth and differentiation of human breast epithelial cells in a three-dimensional (3D co-culture assay. Methods Breast luminal and myoepithelial cells and endothelial cells were isolated from reduction mammoplasties. Primary cells and established normal and malignant breast cell lines were embedded in reconstituted basement membrane in direct co-culture with endothelial cells and by separation of Transwell filters. Morphogenic and phenotypic profiles of co-cultures was evaluated by phase contrast microscopy, immunostaining and confocal microscopy. Results In co-culture, endothelial cells stimulate proliferation of both luminal- and myoepithelial cells. Furthermore, endothelial cells induce a subpopulation of luminal epithelial cells to form large acini/ducts with a large and clear lumen. Endothelial cells also stimulate growth and cloning efficiency of normal and malignant breast epithelial cell lines. Transwell and gradient co-culture studies show that endothelial derived effects are mediated - at least partially - by soluble factors. Conclusion Breast endothelial cells - beside their role in transporting nutrients and oxygen to tissues - are vital component of the epithelial microenvironment in the breast and provide proliferative signals to the normal and malignant breast epithelium. These growth promoting effects of endothelial cells should be taken into consideration in breast cancer biology.
-
Energy Technology Data Exchange (ETDEWEB)
Zhang, Fenxi, E-mail: fxzhang0824@gmail.com [Department of Anatomy, Sanquan College, Xinxiang Medical University, Henan 453003, People' s Republic of China (China); Hong, Yan; Liang, Wenmei [Department of Histology and Embryology, Guiyang Medical University, Guizhou 550004, People' s Republic of China (China); Ren, Tongming [Department of Anatomy, Sanquan College, Xinxiang Medical University, Henan 453003, People' s Republic of China (China); Jing, Suhua [ICU Center, The Third Hospital of Xinxiang Medical University, Henan 453003, People' s Republic of China (China); Lin, Juntang [Stem Cell Center, Xinxiang Medical University, Henan 453003, People' s Republic of China (China)
2012-10-12
Highlights: Black-Right-Pointing-Pointer Co-culture of Sertoli cells (SCs) with human umbilical cord mesenchymal stem cells (UCMSCs). Black-Right-Pointing-Pointer Presence of SCs dramatically increased proliferation and migration of UCMSCs. Black-Right-Pointing-Pointer Presence of SCs stimulated expression of Mdm2, Akt, CDC2, Cyclin D, CXCR4, MAPKs. -- Abstract: Human umbilical cord mesenchymal stem cells (hUCMSCs) have been recently used in transplant therapy. The proliferation and migration of MSCs are the determinants of the efficiency of MSC transplant therapy. Sertoli cells are a kind of 'nurse' cells that support the development of sperm cells. Recent studies show that Sertoli cells promote proliferation of endothelial cells and neural stem cells in co-culture. We hypothesized that co-culture of UCMSCs with Sertoli cells may also promote proliferation and migration of UCMSCs. To examine this hypothesis, we isolated UCMSCs from human cords and Sertoli cells from mouse testes, and co-cultured them using a Transwell system. We found that UCMSCs exhibited strong proliferation ability and potential to differentiate to other cell lineages such as osteocytes and adipocytes. The presence of Sertoli cells in co-culture significantly enhanced the proliferation and migration potential of UCMSCs (P < 0.01). Moreover, these phenotypic changes were accompanied with upregulation of multiple genes involved in cell proliferation and migration including phospho-Akt, Mdm2, phospho-CDC2, Cyclin D1, Cyclin D3 as well as CXCR4, phospho-p44 MAPK and phospho-p38 MAPK. These findings indicate that Sertoli cells boost UCMSC proliferation and migration potential.
-
International Nuclear Information System (INIS)
Yamada, Y; Umegaki, G; Kawashima, T; Nagai, M; Shibata, T; Masuzawa, T; Kimura, T; Kishida, A
2012-01-01
In order to realize a cell culture microdevice actuated by piezoelectric thin films for on-chip regulation of cell functions, this paper reported on a feasibility study by using the microdevice with KOH-etched cavities surrounded by four (111) sidewalls as microchambers in order to introduce cells to be cultured. As a result, the vibration characteristic of the PZT actuator was improved by using an electric field -150 kV/cm at 70 C for 30 min in poling process. A feasibility study on cell culture for delivering mechanical vibratory stimuli to cells revealed the microdevice could be applicable to the culture with actual biological cells. In addition, it was found that O 2 -plasma treated parylene-C process could be applicable for obtaining homogeneous surface of cell culture microdevice.
-
Advancing Our Understanding of Cross-Cultural Issues in Consumer Science and Consumer Psychology
van Herk, H.; Torelli, Carlos J.; van Herk, Hester; Torelli, Carlos J.
2017-01-01
Globalization has resulted in a more complex marketplace. Growing multi-culturalism of consumer markets and increased global competition are pushing marketing scholars to better understand cross-cultural issues in consumer science and consumer psychology. The chapters in this book cover the field to
-
Cell Phones Transform a Science Methods Course
Madden, Lauren
2012-01-01
A science methods instructor intentionally encouraged cell phone use for class work to discover how cell phones can be used as research tools to enhance the content and engage the students. The anecdotal evidence suggested that students who used their smartphones as research tools experienced the science content and pedagogical information…
-
Microfluidic Cell Culture Device
Takayama, Shuichi (Inventor); Cabrera, Lourdes Marcella (Inventor); Heo, Yun Seok (Inventor); Smith, Gary Daniel (Inventor)
2014-01-01
Microfluidic devices for cell culturing and methods for using the same are disclosed. One device includes a substrate and membrane. The substrate includes a reservoir in fluid communication with a passage. A bio-compatible fluid may be added to the reservoir and passage. The reservoir is configured to receive and retain at least a portion of a cell mass. The membrane acts as a barrier to evaporation of the bio-compatible fluid from the passage. A cover fluid may be added to cover the bio-compatible fluid to prevent evaporation of the bio-compatible fluid.
-
Novel culturing platform for brain slices and neuronal cells
DEFF Research Database (Denmark)
Svendsen, Winnie Edith; Al Atraktchi, Fatima Al-Zahraa; Bakmand, Tanya
2015-01-01
In this paper we demonstrate a novel culturing system for brain slices and neuronal cells, which can control the concentration of nutrients and the waste removal from the culture by adjusting the fluid flow within the device. The entire system can be placed in an incubator. The system has been...... tested successfully with brain slices and PC12 cells. The culture substrate can be modified using metal electrodes and/or nanostructures for conducting electrical measurements while culturing and for better mimicking the in vivo conditions....
-
Peptide Hydrogelation and Cell Encapsulation for 3D Culture of MCF-7 Breast Cancer Cells
Sun, Xiuzhi S.; Nguyen, Thu A.
2013-01-01
Three-dimensional (3D) cell culture plays an invaluable role in tumor biology by providing in vivo like microenviroment and responses to therapeutic agents. Among many established 3D scaffolds, hydrogels demonstrate a distinct property as matrics for 3D cell culture. Most of the existing pre-gel solutions are limited under physiological conditions such as undesirable pH or temperature. Here, we report a peptide hydrogel that shows superior physiological properties as an in vitro matrix for 3D cell culture. The 3D matrix can be accomplished by mixing a self-assembling peptide directly with a cell culture medium without any pH or temperature adjustment. Results of dynamic rheological studies showed that this hydrogel can be delivered multiple times via pipetting without permanently destroying the hydrogel architecture, indicating the deformability and remodeling ability of the hydrogel. Human epithelial cancer cells, MCF-7, are encapsulated homogeneously in the hydrogel matrix during hydrogelation. Compared with two-dimensional (2D) monolayer culture, cells residing in the hydrogel matrix grow as tumor-like clusters in 3D formation. Relevant parameters related to cell morphology, survival, proliferation, and apoptosis were analyzed using MCF-7 cells in 3D hydrogels. Interestingly, treatment of cisplatin, an anti-cancer drug, can cause a significant decrease of cell viability of MCF-7 clusters in hydrogels. The responses to cisplatin were dose- and time-dependent, indicating the potential usage of hydrogels for drug testing. Results of confocal microscopy and Western blotting showed that cells isolated from hydrogels are suitable for downstream proteomic analysis. The results provided evidence that this peptide hydrogel is a promising 3D cell culture material for drug testing. PMID:23527204
-
Dynamic cell culture system (7-IML-1)
Cogoli, Augusto
1992-01-01
This experiment is one of the Biorack experiments being flown on the International Microgravity Laboratory 1 (MIL-1) mission as part of an investigation studying cell proliferation and performance in space. One of the objectives of this investigation is to assess the potential benefits of bioprocessing in space with the ultimate goal of developing a bioreactor for continuous cell cultures in space. This experiment will test the operation of an automated culture chamber that was designed for use in a Bioreactor in space. The device to be tested is called the Dynamic Cell Culture System (DCCS). It is a simple device in which media are renewed or chemicals are injected automatically, by means of osmotic pumps. This experiment uses four Type I/O experiment containers. One DCCS unit, which contains a culture chamber with renewal of medium and a second chamber without a medium supply fits in each container. Two DCCS units are maintained under zero gravity conditions during the on-orbit period. The other two units are maintained under 1 gh conditions in a 1 g centrifuge. The schedule for incubator transfer is given.
-
Codrington, Jamila
2014-12-01
Wallace and Brand's framing of culturally responsive science teaching through the lens of critical race theory honors the role of social justice in science education. In this article, I extend the discussion through reflections on the particular learning needs of students from oppressed cultural groups, specifically African Americans. Understanding the political nature of education, I explore the importance of transforming science education so that it has the capacity to provide African American students with tools for their own liberation. I discuss Wallace and Brand's research findings in relation to the goal of liberatory education, and offer ideas for how science educators might push forward this agenda as they strive for culturally responsive teaching with oppressed student groups.
-
Towards a Cultural Psychology of Science
DEFF Research Database (Denmark)
Marco Carre Benzi, David
The present thesis is an enquiry about two distinct but complementary issues: the personal dimension of scientific activity, and the influential role that economists have had during the last decades in Chile. Regarding the former, this work complements existing philosophical, social, and psycholo......The present thesis is an enquiry about two distinct but complementary issues: the personal dimension of scientific activity, and the influential role that economists have had during the last decades in Chile. Regarding the former, this work complements existing philosophical, social......, and psychological studies of science with a cultural psychology perspective. This perspective aims to be sensitive to the personal nature of the scientific activity but also to the cultural conditions in which scientific knowledge is constructed, without subsuming any of these dimensions into the other. At the same...... time, this work offers a novel perspective on the notorious role that economists have had in contemporary Chilean society, a topic that has been mostly addressed as exclusively social and institutional. By focusing on economists’ experiences and views, this thesis shows that, while inserted...
-
Trivalent MDCK cell culture-derived influenza vaccine Optaflu (Novartis Vaccines).
Doroshenko, Alexander; Halperin, Scott A
2009-06-01
Annual influenza epidemics continue to have a considerable impact in both developed and developing countries. Vaccination remains the principal measure to prevent seasonal influenza and reduce associated morbidity and mortality. The WHO recommends using established mammalian cell culture lines as an alternative to egg-based substrates in the manufacture of influenza vaccine. In June 2007, the EMEA approved Optaflu, a Madin Darby canine kidney cell culture-derived influenza vaccine manufactured by Novartis Vaccines. This review examines the advantages and disadvantages of cell culture-based technology for influenza vaccine production, compares immunogenicity and safety data for Optaflu with that of currently marketed conventional egg-based influenza vaccines, and considers the prospects for wider use of cell culture-based influenza vaccines.
-
Fabrication of cell-benign inverse opal hydrogels for three-dimensional cell culture.
Im, Pilseon; Ji, Dong Hwan; Kim, Min Kyung; Kim, Jaeyun
2017-05-15
Inverse opal hydrogels (IOHs) for cell culture were fabricated and optimized using calcium-crosslinked alginate microbeads as sacrificial template and gelatin as a matrix. In contrast to traditional three-dimensional (3D) scaffolds, the gelatin IOHs allowed the utilization of both the macropore surface and inner matrix for cell co-culture. In order to remove templates efficiently for the construction of 3D interconnected macropores and to maintain high cell viability during the template removal process using EDTA solution, various factors in fabrication, including alginate viscosity, alginate concentration, alginate microbeads size, crosslinking calcium concentration, and gelatin network density were investigated. Low viscosity alginate, lower crosslinking calcium ion concentration, and lower concentration of alginate and gelatin were found to obtain high viability of cells encapsulated in the gelatin matrix after removal of the alginate template by EDTA treatment by allowing rapid dissociation and diffusion of alginate polymers. Based on the optimized fabrication conditions, gelatin IOHs showed good potential as a cell co-culture system, applicable to tissue engineering and cancer research. Copyright © 2017 Elsevier Inc. All rights reserved.
-
Yeatts, Andrew B; Choquette, Daniel T; Fisher, John P
2013-02-01
Mesenchymal stem cells (MSCs) are a promising cell source for bone and cartilage tissue engineering as they can be easily isolated from the body and differentiated into osteoblasts and chondrocytes. A cell based tissue engineering strategy using MSCs often involves the culture of these cells on three-dimensional scaffolds; however the size of these scaffolds and the cell population they can support can be restricted in traditional static culture. Thus dynamic culture in bioreactor systems provides a promising means to culture and differentiate MSCs in vitro. This review seeks to characterize key MSC differentiation signaling pathways and provides evidence as to how dynamic culture is augmenting these pathways. Following an overview of dynamic culture systems, discussion will be provided on how these systems can effectively modify and maintain important culture parameters including oxygen content and shear stress. Literature is reviewed for both a highlight of key signaling pathways and evidence for regulation of these signaling pathways via dynamic culture systems. The ability to understand how these culture systems are affecting MSC signaling pathways could lead to a shear or oxygen regime to direct stem cell differentiation. In this way the efficacy of in vitro culture and differentiation of MSCs on three-dimensional scaffolds could be greatly increased. Bioreactor systems have the ability to control many key differentiation stimuli including mechanical stress and oxygen content. The further integration of cell signaling investigations within dynamic culture systems will lead to a quicker realization of the promise of tissue engineering and regenerative medicine. This article is part of a Special Issue entitled Biochemistry of Stem Cells. Copyright © 2012 Elsevier B.V. All rights reserved.
-
Egger, A. E.
2014-12-01
Teachers provide foundational science experiences that spark interest in some students to pursue science and serve as an endpoint for others. For both groups, getting a glimpse into the culture of science is important to their futures as citizens, but this glimpse is not something all teachers are equipped to offer. Explicit instruction in the culture of science is generally not part of college-level science courses; to reach future teachers, it should be incorporated into the curriculum for pre-service teachers. I have incorporated readings from Visionlearning's peer-reviewed, freely available, web-based Process of Science series (http://www.visionlearning.com/en/library/Process-of-Science/49) into my class for pre-service middle-level and secondary science teachers. The readings describe the development of the culture and process of science using deeply embedded examples of scientists and their work. Students reflected on each reading by describing what they learned and something they will use in their future teaching. Responses were graded for thoughtfulness and completeness and later compiled. In general, students with more science courses had a better initial understanding of the culture of science and found the readings engaging stories that explained in more depth what they already knew. However, all students reported learning some fundamental aspects of the culture and nature of science. Most commonly, they learned scientific language, often words with both colloquial and scientific definitions: theory, hypothesis, law, uncertainty, error, confidence. Other learning gains were reported in defining the difference between scientific controversy and social controversy over science, interactions between historical events and the scientific enterprise, how much scientists work in groups and interact at meetings, and the role that funding plays in guiding research. On their own, students struggled to describe explicit ways to incorporate these concepts into their
-
Radiation adaptive response for the growth of cultured glial cells
International Nuclear Information System (INIS)
Suzuki, S.; Miura, Y.; Kano, M.; Toda, T.; Urano, S.
2003-01-01
Full text: To examine the molecular mechanism of radiation adaptive response (RAR) for the growth of cultured glial cells and to investigate the influence of aging on the response, glial cells were cultured from young and aged rats (1 month and 24 months old). RAR for the growth of glial cells conditioned with a low dose of X-rays and subsequently exposed to a high dose of X-rays was examined for cell number and BrdU incorporation. Involvement of the subcellular signaling pathway factors in RAR was investigated using their inhibitors, activators and mutated glial cells. RAR was observed in cells cultured from young rats, but was not in cells from aged rats. The inhibitors of protein kinase C (PKC) and DNA-dependent protein kinase (DNA-PK) or phosphatidylinositol 3-kinase (PI3K) suppressed RAR. The activators of PKC instead of low dose irradiation also caused RAR. Moreover, glial cells cultured from severe combined immunodeficiency (scid) mice (CB-17 scid) and ataxia-telangiectasia (AT) cells from AT patients showed no RAR. These results indicated that PKC, ATM, DNAPK and/or PI3K were involved in RAR for growth and BrdU incorporation of cultured glial cells and RAR decreased with aging. Proteomics data of glial cells exposed to severe stress of H 2 O 2 or X-rays also will be presented in the conference since little or no difference has not been observed with slight stress yet
-
Creating Effective Partnerships in Ecosystem-Based Management: A Culture of Science and Management
Directory of Open Access Journals (Sweden)
Carlie S. Wiener
2011-01-01
Full Text Available An ecosystem-based management research partnership between the Hawai‘i Institute of Marine Biology and Office of National Marine Sanctuaries, specifically with the Northwestern Hawaiian Islands Coral Reef Ecosystem Reserve and, later, the Papahānaumokuākea Marine National Monument, provides a case study to analyze integration of scientific research into management plans through collaborative communications. Ecosystem-based management seeks input from disparate stakeholders and requires effective communication systems for the public, science, and management partners that bypass differences in organizational culture and communication styles. Here, we examine a successful partnership within the framework of ecosystem-based management to survey and evaluate cultural differences, understand what facilitates collaborative communication, highlight factors that impede a successful partnership, and identify areas for improvement. Effective communication has been achieved through an analysis of the organizations cultures and structures to better define communication links. Although specific differences were noted in organization and style, successful integration was accomplished through techniques such as the development of symposia and semiannual reports. This paper will explore the organizational culture analysis and structure evaluation, which are components of a larger study. This science management integration project is an example of how organizational analysis can lead to recommendations for improved communication and integration of science and management.
-
Rakers, Sebastian; Klinger, Matthias; Kruse, Charli; Gebert, Marina
2011-12-01
Here, we report the establishment of a permanent skin cell culture from rainbow trout (Oncorhynchus mykiss). The cells of the fish skin cell culture could be propagated over 60 passages so far. Furthermore, we show for the first time that it is possible to integrate freshly harvested rainbow trout scales into this new fish skin cell culture. We further demonstrated that epithelial cells derived from the scales survived in the artificial micro-environment of surrounding fibroblast-like cells. Also, antibody staining indicated that both cell types proliferated and started to build connections with the other cell type. It seems that it is possible to generate an 'artificial skin' with two different cell types. This could lead to the development of a three-dimensional test system, which might be a better in vitro representative of fish skin in vivo than individual skin cell lines. Copyright © 2011 Elsevier GmbH. All rights reserved.
-
Preparation of labelled lipids by the use of plant cell cultures
International Nuclear Information System (INIS)
Mangold, H.K.
1978-01-01
The preparation of some radioacitvely labelled lipids by the use of plant cell cultures is discussed and further applications of the new method are suggested. Cell suspension cultures of rape (Brassica napus) and soya (Glycine max) have been used for the preparation of lipids labelled with radioisotopes. Radioactive acetic acid as well as various long-chain fatty acids are readily incorporated into the neutral and ionic lipids of plant cell cultures. In addition, 14 C-labelled glycerol, ethanolamine and choline are well utilized by the cells. Randomly labelled lipids have been obtained by incubating cell suspension cultures of rape and soya with [1- 14 C] acetic acid, and uniformly labelled lipids have been isolated from cultures that had been incubated with a mixture of [1- 14 C] acetic acid plus [2- 14 C] acetic acid. The use of techniques of plant cell cultures for the preparation of lipds labelled with stable or radioactive isotopesappears particularly rewarding because the uptake of precursors by the cells and their incorporation into various lipid compounds proceeds rapidly and often quanitatively.This new approach should be useful also for the biosynthesis of lipids whose acyl moieties contain a spn radical, a fluorescent group, or a light-sensitive label. Thus, plant cell cultures constitute valuable new tools for the biosynthetic preparation of a great variety of labelled lipids. (A.G.)
-
International Nuclear Information System (INIS)
Brock, W.A.; Bock, S.P.; Williams, M.; Baker, F.L.
1987-01-01
This study extends the use of the adhesive-tumor-cell culture system to include: over 100 sensitivity measurements at 2.0 Gy; tumorgenicity determinations in nude mice; and flow cytometry of the cells grown in the system. The malignant nature of the growing cells was proved by injecting cells into nude mice. Tumors resulted in 60% of the cases and the histology of each xenograft was similar to that of the human tumor. Flow cytometry was used to obtain DNA histograms of the original cell suspension and of cultures during the two week culture period in order to obtain quantitative information about the growth of aneuploid versus diploid populations. The results thus far demonstrate that 95% of aneuploid populations yield aneuploid growth; of the first 20 cases studied, only one suspension with an aneuploid peak resulted in diploid growth. Of further interest was the observation that it is not unusual for a minor aneuploid population to become the predominate growth fraction after two weeks in culture. These results demonstrate that the adhesive-tumor-cell culture system supports the growth of malignant cells, that multiple cell populations exist in cell suspensions derived from solid tumors, and that differences exist between the radiosensitivity of cells at 2.0 Gy in different histology types
-
Culture conditions defining glioblastoma cells behavior: what is the impact for novel discoveries?
Ledur, Pítia Flores; Onzi, Giovana Ravizzoni; Zong, Hui; Lenz, Guido
2017-09-15
In cancer research, the use of established cell lines has gradually been replaced by primary cell cultures due to their better representation of in vivo cancer cell behaviors. However, a major challenge with primary culture involves the finding of growth conditions that minimize alterations in the biological state of the cells. To ensure reproducibility and translational potentials for research findings, culture conditions need to be chosen so that the cell population in culture best mimics tumor cells in vivo . Glioblastoma (GBM) is one of the most aggressive and heterogeneous tumor types and the GBM research field would certainly benefit from culture conditions that could maintain the original plethora of phenotype of the cells. Here, we review culture media and supplementation options for GBM cultures, the rationale behind their use, and how much those choices affect drug-screening outcomes. We provide an overview of 120 papers that use primary GBM cultures and discuss the current predominant conditions. We also show important primary research data indicating that "mis-cultured" glioma cells can acquire unnatural drug sensitivity, which would have devastating effects for clinical translations. Finally, we propose the concurrent test of four culture conditions to minimize the loss of cell coverage in culture.
-
40 CFR 798.5300 - Detection of gene mutations in somatic cells in culture.
2010-07-01
... cells in culture. 798.5300 Section 798.5300 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY....5300 Detection of gene mutations in somatic cells in culture. (a) Purpose. Mammalian cell culture... selected by resistance to ouabain. (2) Description. Cells in suspension or monolayer culture are exposed to...
-
Using Natural Sciences for Cultural Expansion: The National Socialist Agenda for the Balkans
Directory of Open Access Journals (Sweden)
Maria Zarifi
2008-11-01
Full Text Available This article highlights the political merit natural sciences were awarded under the totalitarian regime of Nazi Germany and their propagandistic role in Hitler's foreign policy agenda for the Balkans, a region which was expected to replace Germany's colonies lost in World War I. It accounts further for the policies and strategies National Socialists used to exert cultural influence on the countries of South-East Europe, namely through a number of institutions with which natural sciences were in one way or another involved in order to promote German culture abroad. The promotion of the German language and, to a certain degree, the Nazi ideology was a precondition for familiarising the Balkan countries with German scientific achievements, which would pave the way for an economic and political infiltration in that region. Therefore, natural sciences, as part of the German intellect, acquired political and economic connotations hidden behind the euphemistic term of cultural policy, designed for this region of geopolitical importance. The article is based almost exclusively on unpublished German records.
-
Prospects for the use of plant cell cultures in food biotechnology.
Davies, Kevin M; Deroles, Simon C
2014-04-01
Plant cell cultures can offer continuous production systems for high-value food and health ingredients, independent of geographical or environmental variations and constraints. Yet despite many improvements in culture technologies, cell line selection, and bioreactor design, there are few commercial successes. This is principally due to the culture yield and market price of food products not being sufficient to cover the plant cell culture production costs. A better understanding of the underpinning biological mechanisms that control the target metabolite biosynthetic pathways may allow the metabolic engineering of cell lines to provide for economically competitive product yields. However, uncertainty around the regulatory and public acceptance of products derived from engineered cell cultures presents a barrier to the uptake of the technology by food product companies. Copyright © 2014 Elsevier Ltd. All rights reserved.
-
Cytotoxicity of extracts of spices to cultured cells.
Unnikrishnan, M C; Kuttan, R
1988-01-01
The cytotoxicity of the extracts from eight different spices used in the Indian diet was determined using Dalton's lymphoma ascites tumor cells and human lymphocytes in vitro and Chinese Hamster Ovary cells and Vero cells in tissue culture. Alcoholic extracts of the spices were found to be more cytotoxic to these cells than their aqueous extracts. Alcoholic extracts of several spices inhibited cell growth at concentrations of 0.2-1 mg/ml in vitro and 0.12-0.3 mg/ml in tissue culture. Ginger, pippali (native to India; also called dried catkins), pepper, and garlic showed the highest activity followed by asafetida, mustard, and horse-gram (native to India). These extracts also inhibited the thymidine uptake into DNA.
-
A Systematic Review: The Next Generation Science Standards and the Increased Cultural Diversity
Asowayan, Alaa A.; Ashreef, Samaar Y.; Omar, Sozan H.
2017-01-01
This systematic review aims to explore the effect of NGSS on students' academic excellence. Specifically, considering increased cultural diversity, it is appropriate to identify student's science-related values, respectful features of teachers' cultural competence, and underlying challenges and detect in what ways these objectives are addressed by…
-
A Novel Counter Sheet-flow Sandwich Cell Culture Device for Mammalian Cell Growth in Space
Sun, Shujin; Gao, Yuxin; Shu, Nanjiang; Tang, Zemei; Tao, Zulai; Long, Mian
2008-08-01
Cell culture and growth in space is crucial to understand the cellular responses under microgravity. The effects of microgravity were coupled with such environment restrictions as medium perfusion, in which the underlying mechanism has been poorly understood. In the present work, a customer-made counter sheet-flow sandwich cell culture device was developed upon a biomechanical concept from fish gill breathing. The sandwich culture unit consists of two side chambers where the medium flow is counter-directional, a central chamber where the cells are cultured, and two porous polycarbonate membranes between side and central chambers. Flow dynamics analysis revealed the symmetrical velocity profile and uniform low shear rate distribution of flowing medium inside the central culture chamber, which promotes sufficient mass transport and nutrient supply for mammalian cell growth. An on-orbit experiment performed on a recovery satellite was used to validate the availability of the device.
-
Enhanced chondrocyte culture and growth on biologically inspired nanofibrous cell culture dishes.
Bhardwaj, Garima; Webster, Thomas J
2016-01-01
Chondral and osteochondral defects affect a large number of people in which treatment options are currently limited. Due to its ability to mimic the natural nanofibrous structure of cartilage, this current in vitro study aimed at introducing a new scaffold, called XanoMatrix™, for cartilage regeneration. In addition, this same scaffold is introduced here as a new substrate onto which to study chondrocyte functions. Current studies on chondrocyte functions are limited due to nonbiologically inspired cell culture substrates. With its polyethylene terephthalate and cellulose acetate composition, good mechanical properties and nanofibrous structure resembling an extracellular matrix, XanoMatrix offers an ideal surface for chondrocyte growth and proliferation. This current study demonstrated that the XanoMatrix scaffolds promote chondrocyte growth and proliferation as compared with the Corning and Falcon surfaces normally used for chondrocyte cell culture. The XanoMatrix scaffolds also have greater hydrophobicity, three-dimensional surface area, and greater tensile strength, making them ideal candidates for alternative treatment options for chondral and osteochondral defects as well as cell culture substrates to study chondrocyte functions.
-
5-Fluorouracil-induced apoptosis in cultured oral cancer cells.
Tong, D; Poot, M; Hu, D; Oda, D
2000-03-01
Chemotherapy is commonly used to treat advanced oral squamous cell carcinoma (SCC) and is known to kill cancer cells through apoptosis. Our hypothesis states that 5-fluorouracil (5FU) also kills cultured oral epithelial cells through programmed cell death or apoptosis. Cultured oral cancer cells were exposed to an optimum dose of 20 mg/ml of 5FU. Cells were analyzed for changes in cell cycle distribution and induction of cell death including apoptosis. Normal control, human papilloma virus-immortalized (PP), ATCC SCC cell line (CA1) and two primary oral SCC cell lines (CA3 and -4) were studied. Inhibition of apoptosis by a pan-caspase inhibitor was used. SYTO 11 flow cytometry showed increased apoptosis in all 5FU-treated cell cultures compared to untreated controls. The results show biological variation in apoptotic response. CA1 had the lowest apoptotic rate of the cancer cell lines at 1.5%. Next lowest was CA3, followed by CA4 and PP. In addition, alteration in the G1 and S phase fractions were found. Untreated CA1 showed 28% G1, 53% S compared to 43% G1, and 40% S of treated. We investigated the pathway of apoptosis using the pan-caspase inhibitor IDN-1529 by methylthiazolyl diphenyl tetrazolium bromide (MTT) colorimetric analysis. Results showed mild inhibition of cell death when cells were incubated with 50 microM IDN-1529 for 24 h. This suggests a probable caspase-dependent apoptotic pathway. In conclusion, our data suggest that 5FU induces oral cancer cell death through apoptosis and that biological variation exists between normal and cancer cells and between different types of cancer cells themselves. Our data indicate that cultures of a useful in vitro model for chemosensitivity assays are possible. Our results also suggest a caspase-dependent pathway for chemocytotoxicity in oral SCC.
-
2011-09-19
... Denver Museum of Nature & Science, Denver, CO, that meets the definition of an object of cultural... Cultural Item: Denver Museum of Nature and Science, Denver, CO AGENCY: National Park Service, Interior. ACTION: Notice. SUMMARY: The Denver Museum of Nature & Science, in consultation with the appropriate...
-
DEFF Research Database (Denmark)
Petronis, Sarunas; Stangegaard, Michael; Christensen, C.
2006-01-01
Modern microfabrication and microfluidic technologies offer new opportunities in the design and fabrication of miniaturized cell culture systems for online monitoring of living cells. We used laser micromachining and thermal bonding to fabricate an optically transparent, low-cost polymeric chip...... for long-term online cell culture observation under controlled conditions. The chip incorporated a microfluidic flow equalization system, assuring uniform perfusion of the cell culture media throughout the cell culture chamber. The integrated indium-tin-oxide heater and miniature temperature probe linked....... HeLa cells were cultured for up to 2 weeks within the cell culture chip and monitored using a time-lapse video recording microscopy setup. Cell attachment and spreading was observed during the first 10-20 h (lag phase). After approximately 20 h, cell growth gained exponential character...
-
Science or liberal arts? Cultural capital and college major choice in China.
Hu, Anning; Wu, Xiaogang
2017-12-19
Previous studies on major East Asian societies such as Japan and Korea generally fail to find a strong effect of cultural capital in educational inequality, partly due to the characteristic extreme focus on standardized test and curriculum. This study shifts attention to the horizontal stratification of education by investigating the association between family background, cultural capital, and college major choice in contemporary China. Based on analysis of data from the Beijing College Students Panel Survey (BCSPS), we found that, on average, cultural capital significantly mediates the relationship between family background and college major preference. Those with greater endowment of cultural capital are more likely to come from socio-economically advantaged families, and, at the same time, demonstrate a stronger propensity to major in liberal arts fields rather than science, technology, engineering and mathematics (STEM) fields. Further analyses reveal that the association between cultural capital and academic field choice comes into being by way of performance in the Chinese test in the national college entrance examination and of the non-cognitive dispositions, such as self-efficacy and self-esteem. Our findings better our understanding of formation of the horizontal stratification of higher education. © London School of Economics and Political Science 2017.
-
Haraguchi, Yuji; Matsuura, Katsuhisa; Shimizu, Tatsuya; Yamato, Masayuki; Okano, Teruo
2015-12-01
In this study, a simple three-dimensional (3D) suspension culture method for the expansion and cardiac differentiation of human induced pluripotent stem cells (hiPSCs) is reported. The culture methods were easily adapted from two-dimensional (2D) to 3D culture without any additional manipulations. When hiPSCs were directly applied to 3D culture from 2D in a single-cell suspension, only a few aggregated cells were observed. However, after 3 days, culture of the small hiPSC aggregates in a spinner flask at the optimal agitation rate created aggregates which were capable of cell passages from the single-cell suspension. Cell numbers increased to approximately 10-fold after 12 days of culture. The undifferentiated state of expanded hiPSCs was confirmed by flow cytometry, immunocytochemistry and quantitative RT-PCR, and the hiPSCs differentiated into three germ layers. When the hiPSCs were subsequently cultured in a flask using cardiac differentiation medium, expression of cardiac cell-specific genes and beating cardiomyocytes were observed. Furthermore, the culture of hiPSCs on Matrigel-coated dishes with serum-free medium containing activin A, BMP4 and FGF-2 enabled it to generate robust spontaneous beating cardiomyocytes and these cells expressed several cardiac cell-related genes, including HCN4, MLC-2a and MLC-2v. This suggests that the expanded hiPSCs might maintain the potential to differentiate into several types of cardiomyocytes, including pacemakers. Moreover, when cardiac cell sheets were fabricated using differentiated cardiomyocytes, they beat spontaneously and synchronously, indicating electrically communicative tissue. This simple culture system might enable the generation of sufficient amounts of beating cardiomyocytes for use in cardiac regenerative medicine and tissue engineering. Copyright © 2013 John Wiley & Sons, Ltd.
-
DEFF Research Database (Denmark)
Hellsten, Ylva; Frandsen, Ulrik
1997-01-01
1. The present study examined the capacity for adenosine formation, uptake and metabolism in contracting primary rat muscle cells and in microvascular endothelial cells in culture. 2. Strong and moderate electrical simulation of skeletal muscle cells led to a significantly greater increase....... 3. Addition of microvascular endothelial cells to the cultured skeletal muscle cells enhanced the contraction-induced accumulation of extracellular adenosine (P Skeletal muscle cells were...... in the extracellular adenosine concentration (421 +/- 91 and 235 +/- 30 nmol (g protein)-1, respectively; P muscle cells (161 +/- 20 nmol (g protein)-1). The ATP concentration was lower (18%; P contracted, but not in the moderately contracted muscle cells...
-
Yang, R; Elankumaran, Y; Hijjawi, N; Ryan, U
2015-06-01
A cell-free culture system for Cryptosporidium parvum was analysed using scanning electron microscopy (SEM) to characterise life cycle stages and compare gene expression in cell-free culture and cell culture using HCT-8 cells. Cryptosporidium parvum samples were harvested at 2 h, 8 h, 14 h, 26 h, 50 h, 74 h, 98 h, 122 h and 170 h, chemically fixed and specimens were observed using a Zeiss 1555 scanning electron microscope. The presence of sporozoites, trophozoites and type I merozoites were identified by SEM. Gene expression in cell culture and cell-free culture was studied using reverse transcriptase quantitative PCR (RT-qPCR) of the sporozoite surface antigen protein (cp15), the glycoprotein 900 (gp900), the Cryptosporidium oocyst wall protein (COWP) and 18S ribosomal RNA (rRNA) genes in both cell free and conventional cell culture. In cell culture, cp15 expression peaked at 74 h, gp900 expression peaked at 74 h and 98 h and COWP expression peaked at 50 h. In cell-free culture, CP15 expression peaked at 98 h, gp900 expression peaked at 74 h and COWP expression peaked at 122 h. The present study is the first to compare gene expression of C. parvum in cell culture and cell-free culture and to characterise life cycle stages of C. parvum in cell-free culture using SEM. Findings from this study showed that gene expression patterns in cell culture and cell-free culture were similar but in cell-free culture, gene expression was delayed for CP15 and COWP in cell free culture compared with the cell culture system and was lower. Although three life cycle stageswere conclusively identified, improvements in SEM methodology should lead to the detection of more life cycle stages. Copyright © 2015 Elsevier Inc. All rights reserved.
-
Yongky, Andrew; Lee, Jongchan; Le, Tung; Mulukutla, Bhanu Chandra; Daoutidis, Prodromos; Hu, Wei-Shou
2015-07-01
Continuous culture for the production of biopharmaceutical proteins offers the possibility of steady state operations and thus more consistent product quality and increased productivity. Under some conditions, multiplicity of steady states has been observed in continuous cultures of mammalian cells, wherein with the same dilution rate and feed nutrient composition, steady states with very different cell and product concentrations may be reached. At those different steady states, cells may exhibit a high glycolysis flux with high lactate production and low cell concentration, or a low glycolysis flux with low lactate and high cell concentration. These different steady states, with different cell concentration, also have different productivity. Developing a mechanistic understanding of the occurrence of steady state multiplicity and devising a strategy to steer the culture toward the desired steady state is critical. We establish a multi-scale kinetic model that integrates a mechanistic intracellular metabolic model and cell growth model in a continuous bioreactor. We show that steady state multiplicity exists in a range of dilution rate in continuous culture as a result of the bistable behavior in glycolysis. The insights from the model were used to devise strategies to guide the culture to the desired steady state in the multiple steady state region. The model provides a guideline principle in the design of continuous culture processes of mammalian cells. © 2015 Wiley Periodicals, Inc.
-
Restoring local spiritual and cultural values in science education: The case of Ethiopia
Faris, Solomon Belay
It has been repeatedly observed that home and local context matter in the education of children. A smooth transition between home and classroom prepares children for enjoyable and meaningful life-long learning. Knowledge building in children is influenced by previous experience, values, beliefs and sociocultural factors associated with community. Against this theoretical background, the thesis examined the integration of local spiritual and cultural values to improve science education in Ethiopia. This autoethnographic research used in-depth interviews, supplementary observations and focus group discussion and my biography to identify the perception and practice of common and unique spiritual and cultural values. The study examined whether these values were included and/or excluded in the school curriculum and explored the possibilities for incorporating values in science education and the anticipated tensions resulting from their inclusion. Students, science teachers, parents, employers, curriculum experts, policymakers, elders, and religious leaders participated in the research, conducted in a randomly selected secondary school in Addis Ababa. The sampling followed a kind of snowball method, with a total of twenty key informants participating in interviews, fifteen classroom observations, and one focus group discussion. The data collection aimed at generating stories, which underlie the auto-ethnography methodology. Findings indicated that belief in and fear of God animated and sustained the Ethiopian way of life. Although spiritual teachings derived from sacred writings were the initial foundation for Ethiopian cultural norms, the two merged together later, creating a mosaic pervading every aspect of life in Ethiopia. Education was sustained on this merger of spiritual and cultural norms and values. It was also shown that the now century-old system of formal education did not incorporate those local spiritual and cultural values. Current science education also
-
Miniature Bioreactor System for Long-Term Cell Culture
Gonda, Steve R.; Kleis, Stanley J.; Geffert, Sandara K.
2010-01-01
A prototype miniature bioreactor system is designed to serve as a laboratory benchtop cell-culturing system that minimizes the need for relatively expensive equipment and reagents and can be operated under computer control, thereby reducing the time and effort required of human investigators and reducing uncertainty in results. The system includes a bioreactor, a fluid-handling subsystem, a chamber wherein the bioreactor is maintained in a controlled atmosphere at a controlled temperature, and associated control subsystems. The system can be used to culture both anchorage-dependent and suspension cells, which can be either prokaryotic or eukaryotic. Cells can be cultured for extended periods of time in this system, and samples of cells can be extracted and analyzed at specified intervals. By integrating this system with one or more microanalytical instrument(s), one can construct a complete automated analytical system that can be tailored to perform one or more of a large variety of assays.
-
Brom-de-Luna, Joao Gatto; Canesin, Heloísa Siqueira; Wright, Gus; Hinrichs, Katrin
2018-03-01
Nuclear transfer using somatic cells from frozen semen (FzSC) would allow cloning of animals for which no other genetic material is available. Horses are one of the few species for which cloning is commercially feasible; despite this, there is no information available on the culture of equine FzSC. After preliminary trials on equine FzSC, recovered by density-gradient centrifugation, resulted in no growth, we hypothesized that sperm in the culture system negatively affected cell proliferation. Therefore, we evaluated culture of FzSC isolated using fluorescence-assisted cell sorting. In Exp. 1, sperm were labeled using antibodies to a sperm-specific antigen, SP17, and unlabeled cells were collected. This resulted in high sperm contamination. In Exp. 2, FzSC were labeled using an anti-MHC class I antibody. This resulted in an essentially pure population of FzSC, 13-25% of which were nucleated. Culture yielded no proliferation in any of nine replicates. In Exp. 3, 5 × 10 3 viable fresh, cultured horse fibroblasts were added to the frozen-thawed, washed semen, then this suspension was labeled and sorted as for Exp. 2. The enriched population had a mean of five sperm per recovered somatic cell; culture yielded formation of monolayers. In conclusion, an essentially pure population of equine FzSC could be obtained using sorting for presence of MHC class I antigens. No equine FzSC grew in culture; however, the proliferation of fibroblasts subjected to the same processing demonstrated that the labeling and sorting methods, and the presence of few sperm in culture, were compatible with cell viability. Copyright © 2017 Elsevier B.V. All rights reserved.
-
Wang, Ya-Ling; Liang, Jyh-Chong; Tsai, Chin-Chung
2018-01-01
Science learning self-efficacy could be regarded as a multi-factor belief which comprises different aspects such as cognitive skills, practical work, and everyday application. However, few studies have investigated the relationships among these factors that compose science learning self-efficacy. Also, culture may play an important role in…
-
Exploring the Impact of Culture- and Language-Influenced Physics on Science Attitude Enhancement
Morales, Marie Paz E.
2016-01-01
"Culture," a set of principles that trace and familiarize human beings within their existential realities, may provide an invisible lens through which reality could be discerned. Critically explored in this study is how culture- and language-sensitive curriculum materials in physics improve Pangasinan learners' attitude toward science.…
-
Effects of different feeder layers on culture of bovine embryonic stem cell-like cells in vitro.
Cong, Shan; Cao, Guifang; Liu, Dongjun
2014-12-01
To find a suitable feeder layer is important for successful culture conditions of bovine embryonic stem cell-like cells. In this study, expression of pluripotency-related genes OCT4, SOX2 and NANOG in bovine embryonic stem cell-like cells on mouse embryonic fibroblast feeder layers at 1-5 passages were monitored in order to identify the possible reason that bovine embryonic stem cell-like cells could not continue growth and passage. Here, we developed two novel feeder layers, mixed embryonic fibroblast feeder layers of mouse and bovine embryonic fibroblast at different ratios and sources including mouse fibroblast cell lines. The bovine embryonic stem cell-like cells generated in our study displayed typical stem cell morphology and expressed specific markers such as OCT4, stage-specific embryonic antigen 1 and 4, alkaline phosphatase, SOX2, and NANOG mRNA levels. When feeder layers and cell growth factors were removed, the bovine embryonic stem cell-like cells formed embryoid bodies in a suspension culture. Furthermore, we compared the expression of the pluripotent markers during bovine embryonic stem cell-like cell in culture on mixed embryonic fibroblast feeder layers, including mouse fibroblast cell lines feeder layers and mouse embryonic fibroblast feeder layers by real-time quantitative polymerase chain reaction. Results suggested that mixed embryonic fibroblast and sources including mouse fibroblast cell lines feeder layers were more suitable for long-term culture and growth of bovine embryonic stem cell-like cells than mouse embryonic fibroblast feeder layers. The findings may provide useful experimental data for the establishment of an appropriate culture system for bovine embryonic stem cell lines.
-
Kieninger, J; Aravindalochanan, K; Sandvik, J A; Pettersen, E O; Urban, G A
2014-04-01
Here we present an application, in two tumour cell lines, based on the Sensing Cell Culture Flask system as a cell culture monitoring tool for pericellular oxygen sensing. T-47D (human breast cancer) and T98G (human brain cancer) cells were cultured either in atmospheric air or in a glove-box set at 4% oxygen, in both cases with 5% CO2 in the gas phase. Pericellular oxygen tension was measured with the help of an integrated sensor chip comprising oxygen sensor arrays. Obtained results illustrate variation of pericellular oxygen tension in attached cells covered by stagnant medium. Independent of incubation conditions, low pericellular oxygen concentration levels, usually associated with hypoxia, were found in dense cell cultures. Respiration alone brought pericellular oxygen concentration down to levels which could activate hypoxia-sensing regulatory processes in cultures believed to be aerobic. Cells in culture believed to experience conditions of mild hypoxia may, in reality, experience severe hypoxia. This would lead to incorrect assumptions and suggests that pericellular oxygen concentration readings are of great importance to obtain reproducible results when dealing with hypoxic and normoxic (aerobic) incubation conditions. The Sensing Cell Culture Flask system allows continuous monitoring of pericellular oxygen concentration with outstanding long-term stability and no need for recalibration during cell culture experiments. The sensor is integrated into the flask bottom, thus in direct contact with attached cells. No additional equipment needs to be inserted into the flask during culturing. Transparency of the electrochemical sensor chip allows optical inspection of cells attached on top of the sensor. © 2014 John Wiley & Sons Ltd.
-
PDMS/glass microfluidic cell culture system for cytotoxicity tests and cells passage
DEFF Research Database (Denmark)
Ziolkowska, K.; Jedrych, E.; Kwapiszewski, R.
2010-01-01
In this paper, hybrid (PDMS/glass) microfluidic cell culture system (MCCS) integrated with the concentration gradient generator (CGG) is presented. PDMS gas permeability enabled cells' respiration in the fabricated microdevices and excellent glass hydrophilicity allowed successful cells' seeding...
-
Lethal graft-versus-host disease: modification with allogeneic cultured donor cells
International Nuclear Information System (INIS)
Mauch, P.; Lipton, J.M.; Hamilton, B.; Obbagy, J.; Kudisch, M.; Nathan, D.; Hellman, S.
1984-01-01
The use of the bone marrow culture technique was studied as a means to prepare donor marrow for bone marrow transplantation to avoid lethal graft-versus-host disease (GVHD). Preliminary experiments demonstrated the rapid loss of theta-positive cells in such cultures, so that theta-positive cells were not detected after 6 days. Initial experiments in C3H/HeJ (H-2k, Hbbd) recipients prepared with 900 rad demonstrated improved survival when 3-day cultured C57BL/6 (H-2b, Hbbs) donor cells were used in place of hind limb marrow for transplantation. However, hemoglobin typing of recipient animals revealed only short-term donor engraftment, with competitive repopulation of recipient marrow occurring. Subsequent experiments were done in 1,200-rad prepared recipients, with long-term donor engraftment demonstrated. The majority of 1,200-rad prepared animals receiving cultured allogeneic cells died of GVHD, but animals receiving 28-day cultured cells had an improved 90-day survival and a delay in GVHD development over animals receiving hind limb marrow or marrow from shorter times in culture. In addition, animals receiving anti-theta-treated, 3-day nonadherent cells had an improved survival (44%) over animals receiving anti-theta-treated hind limb marrow (20%). These experiments demonstrate modest benefit for the use of cultured cells in bone marrow transplantation across major H-2 histocompatibility complex differences
-
Culturally relevant science: An approach to math science education for Hispanics
Energy Technology Data Exchange (ETDEWEB)
Montellano, B.O. de
1996-11-14
This report describes later stages of a program to develop culturally relevant science and math programs for Hispanic students. Part of this effort was follow-up with 17 teachers who participated in early stages of the program. Response was not very good. Included with the report is a first draft effort for curriculum materials which could be used as is in such a teaching effort. Several of the participating teachers were invited to a writing workshop, where lesson plans were drafted, and critiqued and following rework are listed in this publication. Further work needs to be completed and is ongoing.
-
International Nuclear Information System (INIS)
Rettig, W.F.; Garin-Chesa, P.; Beresford, H.R.; Oettgen, H.F.; Melamed, M.R.; Old, L.J.
1988-01-01
Normal differentiation and malignant transformation of human cells are characterized by specific changes in surface antigen phenotype. In the present study, the authors have defined six cell-surface antigens of human sarcomas and normal mesenchymal cells, by using mixed hemadsorption assays and immunochemical methods for the analysis of cultured cells and immunohistochemical staining for the analysis of normal tissues and > 200 tumor specimens. Differential patterns of F19, F24, G171, G253, S5, and Thy-1 antigen expression were found to characterize (i) subsets of cultured sarcoma cell lines, (ii) cultured fibroblasts derived from various organs, (iii) normal resting and activated mesenchymal tissues, and (iv) sarcoma and nonmesenchymal tumor tissues. These results provide a basic surface antigenic map for cultured mesenchymal cells and mesenchymal tissues and permit the classification of human sarcomas according to their antigenic phenotypes
-
Feeder Cell Type Affects the Growth of In Vitro Cultured Bovine Trophoblast Cells
Directory of Open Access Journals (Sweden)
Islam M. Saadeldin
2017-01-01
Full Text Available Trophectoderm cells are the foremost embryonic cells to differentiate with prospective stem-cell properties. In the current study, we aimed at improving the current approach for trophoblast culture by using granulosa cells as feeders. Porcine granulosa cells (PGCs compared to the conventional mouse embryonic fibroblasts (MEFs were used to grow trophectoderm cells from hatched bovine blastocysts. Isolated trophectoderm cells were monitored and displayed characteristic epithelial/cuboidal morphology. The isolated trophectoderm cells expressed mRNA of homeobox protein (CDX2, cytokeratin-8 (KRT8, and interferon tau (IFNT. The expression level was higher on PGCs compared to MEFs throughout the study. In addition, primary trophectoderm cell colonies grew faster on PGCs, with a doubling time of approximately 48 hrs, compared to MEFs. PGCs feeders produced a fair amount of 17β-estradiol and progesterone. We speculated that the supplementation of sex steroids and still-unknown factors during the trophoblasts coculture on PGCs have helped to have better trophectoderm cell’s growth than on MEFs. This is the first time to use PGCs as feeders to culture trophectoderm cells and it proved superior to MEFs. We propose PGCs as alternative feeders for long-term culture of bovine trophectoderm cells. This model will potentially benefit studies on the early trophoblast and embryonic development in bovines.
-
Sada, Takao; Fujigaya, Tsuyohiko; Niidome, Yasuro; Nakazawa, Kohji; Nakashima, Naotoshi
2011-06-28
Unique near-IR optical properties of single-walled carbon nanotube (SWNTs) are of interest in many biological applications. Here we describe the selective cell detachment and collection from an SWNT-coated cell-culture dish triggered by near-IR pulse laser irradiation. First, HeLa cells were cultured on an SWNT-coated dish prepared by a spraying of an aqueous SWNT dispersion on a glass dish. The SWNT-coated dish was found to show a good cell adhesion behavior as well as a cellular proliferation rate similar to a conventional glass dish. We discovered, by near-IR pulse laser irradiation (at the laser power over 25 mW) to the cell under optical microscopic observation, a quick single-cell detachment from the SWNT-coated surface. Shockwave generation from the irradiated SWNTs is expected to play an important role for the cell detachment. Moreover, we have succeeded in catapulting the target single cell from the cultured medium when the depth of the medium was below 150 μm and the laser power was stronger than 40 mW. The captured cell maintained its original shape. The retention of the genetic information of the cell was confirmed by the polymerase chain reaction (PCR) technique. A target single-cell collection from a culture medium under optical microscopic observation is significant in wide fields of single-cell studies in biological areas.
-
Establishment of primary bovine intestinal epithelial cell culture and clone method.
Zhan, Kang; Lin, Miao; Liu, Ming-Mei; Sui, Yang-Nan; Zhao, Guo-Qi
2017-01-01
The aim of this study was to establish bovine intestinal epithelial cell (BIEC) line and provide a novel clone cell method. Although various strategies of bovine cell culture and clone techniques have been reported, these methods remain not established. Here, we culture successfully primary BIECs and establish a novel clone cell method. Our result showed that BIECs could be successfully cultured and passaged about generation 5. These cellular aggregates and clusters were adherent loosely at day 2 of culture. Cell aggregates and clusters start to proliferate after approximately 4 d. The BIECs showed positive reaction against cytokeratin 18, E-cadherin, and characteristics of epithelial-like morphology. In addition, the fatty acid-binding proteins (FABPs), villin, and intestinal peptidase (IP) band were positive in BIECs. Our results suggest that the establishment of culturing and clone BIEC methods will apply to isolate and clone other primary cells. These BIECs could therefore contribute to the study of bovine intestinal nutrient absorption and regulation, immune regulation, and the pathogenesis of the bovine intestinal disease, which will provide intestinal cell model in vitro.
-
3D Cell Culture in a Self-Assembled Nanofiber Environment.
Directory of Open Access Journals (Sweden)
Yi Wen Chai
Full Text Available The development and utilization of three-dimensional cell culture platforms has been gaining more traction. Three-dimensional culture platforms are capable of mimicking in vivo microenvironments, which provide greater physiological relevance in comparison to conventional two-dimensional cultures. The majority of three-dimensional culture platforms are challenged by the lack of cell attachment, long polymerization times, and inclusion of undefined xenobiotics, and cytotoxic cross-linkers. In this study, we review the use of a highly defined material composed of naturally occurring compounds, hyaluronic acid and chitosan, known as Cell-Mate3DTM. Moreover, we provide an original measurement of Young's modulus using a uniaxial unconfined compression method to elucidate the difference in microenvironment rigidity for acellular and cellular conditions. When hydrated into a tissue-like hybrid hydrocolloid/hydrogel, Cell-Mate3DTM is a highly versatile three-dimensional culture platform that enables downstream applications such as flow cytometry, immunostaining, histological staining, and functional studies to be applied with relative ease.
-
The Two Cultures of Electricity: Between Entertainment and Edification in Victorian Science
Morus, Iwan Rhys
2007-06-01
Reviewing Fleeming Jenkin's Electricity and Magnetism in Nature in 1873 an anonymous reviewer (probably James Clerk Maxwell) remarked that "at the present time there are two sciences of electricity — one that of the lecture-room and the popular treatise; the other that of the testing-office and the engineer's specifications." In this paper I want to look behind Maxwell's remark and examine the relationship between the "two sciences" of electricity during the third quarter or so of the 19th century. In particular I want to look at them in terms of their instrumental technologies. How did apparatus travel between the lecture-room or exhibition-hall and the testing-office or the laboratory? How did skills cross between these different spaces? How did the earlier Victorian culture of electricity as "entertainment and edification" become transformed into late 19th century metrological culture? How did these cultures overlap and how did they differ?
-
Energy Technology Data Exchange (ETDEWEB)
Montellano, B.O. de
1996-11-14
This progress report summarizes results of a teacher workshop. A letter sent to 17 teachers who had participated in the workshop requested a report of any activities undertaken and copies of lesson plans and materials developed. Only nine responses were received, and not all of them demonstrated a satisfactory level of activity. Teachers who submitted materials showing the most promise were invited to participate in the Summer Writing Workshop. A partial first draft of a companion volume for the teacher`s manual was written which provides a rationale for culturally relevant science and presents the cultural and scientific background needed. The outline of the book is presented in Appendix 1. Appendix 2 is a sample chapter from the book.
-
The effects of professional development on science teaching practices and classroom culture
Supovitz, Jonathan A.; Turner, Herbert M.
2000-11-01
The current science education reform movement emphasizes the importance of professional development as a means of improving student science achievement. Reformers have developed a vision for professional development based upon intensive and sustained training around concrete tasks that is focused on subject-matter knowledge, connected to specific standards for student performance, and embedded in a systemic context. Using data from a National Science Foundation Teacher Enhancement program called the Local Systemic Change initiative, this study employs hierarchical linear modeling to examine the relationship between professional development and the reformers' vision of teaching practice. The findings indicate that the quantity of professional development in which teachers participate is strongly linked with both inquiry-based teaching practice and investigative classroom culture. At the individual level, teachers' content preparation also has a powerful influence on teaching practice and classroom culture. At the school level, school socioeconomic status was found to influence practice more substantially than either principal supportiveness or available resources.
-
Meier, Lori T.
2012-11-01
This ethnographic case study investigated the science practices of teachers at one public elementary magnet school in light of how school culture influenced science curriculum design and instruction. The purpose of the study was to address how school culture impacted the school's overall treatment of science as a viable content area. Key informant teachers were interviewed to explore their personal beliefs and values, teaching, access to materials, and views of the adopted integrated thematic curriculum model and magnet structure. The resulting data, triangulated with informal observation and artifact collection, were analyzed using a theoretical framework that emphasized five interdependent school culture indicators (values, beliefs, practices, materials, and problems). Findings suggest that the school's culture adversely influenced the treatment of science.
-
Ledur, Pítia F; Liu, Chong; He, Hua; Harris, Alexandra R; Minussi, Darlan C; Zhou, Hai-Yan; Shaffrey, Mark E; Asthagiri, Ashok; Lopes, Maria Beatriz S; Schiff, David; Lu, Yi-Cheng; Mandell, James W; Lenz, Guido; Zong, Hui
2016-10-01
Cell culture plays a pivotal role in cancer research. However, culture-induced changes in biological properties of tumor cells profoundly affect research reproducibility and translational potential. Establishing culture conditions tailored to the cancer cell of origin could resolve this problem. For glioma research, it has been previously shown that replacing serum with defined growth factors for neural stem cells (NSCs) greatly improved the retention of gene expression profile and tumorigenicity. However, among all molecular subtypes of glioma, our laboratory and others have previously shown that the oligodendrocyte precursor cell (OPC) rather than the NSC serves as the cell of origin for the proneural subtype, raising questions regarding the suitability of NSC-tailored media for culturing proneural glioma cells. OPC-originated mouse glioma cells were cultured in conditions for normal OPCs or NSCs, respectively, for multiple passages. Gene expression profiles, morphologies, tumorigenicity, and drug responsiveness of cultured cells were examined in comparison with freshly isolated tumor cells. OPC media-cultured glioma cells maintained tumorigenicity, gene expression profiles, and morphologies similar to freshly isolated tumor cells. In contrast, NSC-media cultured glioma cells gradually lost their OPC features and most tumor-initiating ability and acquired heightened sensitivity to temozolomide. To improve experimental reproducibility and translational potential of glioma research, it is important to identify the cell of origin, and subsequently apply this knowledge to establish culture conditions that allow the retention of native properties of tumor cells. © The Author(s) 2016. Published by Oxford University Press on behalf of the Society for Neuro-Oncology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.
-
Breast Cancer Stem Cell Culture and Enrichment Using Poly(ε-Caprolactone Scaffolds
Directory of Open Access Journals (Sweden)
Sònia Palomeras
2016-04-01
Full Text Available The cancer stem cell (CSC population displays self-renewal capabilities, resistance to conventional therapies, and a tendency to post-treatment recurrence. Increasing knowledge about CSCs’ phenotype and functions is needed to investigate new therapeutic strategies against the CSC population. Here, poly(ε-caprolactone (PCL, a biocompatible polymer free of toxic dye, has been used to fabricate scaffolds, solid structures suitable for 3D cancer cell culture. It has been reported that scaffold cell culture enhances the CSCs population. A RepRap BCN3D+ printer and 3 mm PCL wire were used to fabricate circular scaffolds. PCL design and fabrication parameters were first determined and then optimized considering several measurable variables of the resulting scaffolds. MCF7 breast carcinoma cell line was used to assess scaffolds adequacy for 3D cell culture. To evaluate CSC enrichment, the Mammosphere Forming Index (MFI was performed in 2D and 3D MCF7 cultures. Results showed that the 60° scaffolds were more suitable for 3D culture than the 45° and 90° ones. Moreover, 3D culture experiments, in adherent and non-adherent conditions, showed a significant increase in MFI compared to 2D cultures (control. Thus, 3D cell culture with PCL scaffolds could be useful to improve cancer cell culture and enrich the CSCs population.
-
Cheryan, Sapna; Master, Allison; Meltzoff, Andrew N.
2015-01-01
Despite having made significant inroads into many traditionally male-dominated fields (e.g., biology, chemistry), women continue to be underrepresented in computer science and engineering. We propose that students’ stereotypes about the culture of these fields—including the kind of people, the work involved, and the values of the field—steer girls away from choosing to enter them. Computer science and engineering are stereotyped in modern American culture as male-oriented fields that involve ...
-
Impact of cell culture process changes on endogenous retrovirus expression.
Brorson, Kurt; De Wit, Christina; Hamilton, Elizabeth; Mustafa, Mehnaz; Swann, Patrick G; Kiss, Robert; Taticek, Ron; Polastri, Gian; Stein, Kathryn E; Xu, Yuan
2002-11-05
Cell culture process changes (e.g., changes in scale, medium formulation, operational conditions) and cell line changes are common during the development life cycle of a therapeutic protein. To ensure that the impact of such process changes on product quality and safety is minimal, it is standard practice to compare critical product quality and safety attributes before and after the changes. One potential concern introduced by cell culture process improvements is the possibility of increased endogenous retrovirus expression to a level above the clearance capability of the subsequent purification process. To address this, retrovirus expression was measured in scaled down and full production scaled Chinese hamster ovary (CHO) cell cultures of four monoclonal antibodies and one recombinant protein before and after process changes. Two highly sensitive, quantitative (Q)-PCR-based assays were used to measure endogenous retroviruses. It is shown that cell culture process changes that primarily alter media components, nutrient feed volume, seed density, cell bank source (i.e., master cell bank vs. working cell bank), and vial size, or culture scale, singly or in combination, do not impact the rate of retrovirus expression to an extent greater than the variability of the Q-PCR assays (0.2-0.5 log(10)). Cell culture changes that significantly alter the metabolic state of the cells and/or rates of protein expression (e.g., pH and temperature shifts, NaButyrate addition) measurably impact the rate of retrovirus synthesis (up to 2 log(10)). The greatest degree of variation in endogenous retrovirus expression was observed between individual cell lines (up to 3 log(10)). These data support the practice of measuring endogenous retrovirus output for each new cell line introduced into manufacturing or after process changes that significantly increase product-specific productivity or alter the metabolic state, but suggest that reassessment of retrovirus expression after other
-
Omlor, G W; Nerlich, A G; Tirlapur, U K; Urban, J P; Guehring, T
2014-12-01
Embryonic notochordal disc nucleus cells (NC) have been identified to protect disc tissue against disc degeneration but in human beings NC phenotype gets lost with aging and the pathophysiological mechanisms are poorly understood. NC may stimulate other cells via soluble factors, and NC-conditioned medium can be used to stimulate matrix production of other disc cells and mesenchymal stem cells and thus may be of special interest for biological disc repair. As this stimulatory effect is associated with the NC phenotype, we investigated how cell morphology and gene-expression of the NC phenotype changes with time in 3D-cell culture. NC and inner annulus chondrocyte-like cells (CLC) from immature pigtails (freshly isolated cells/tissue, 3D-alginate beads, 3D-clusters) were cultured for up to 16 days under normoxia and hypoxia. Protein-expression was analysed by immunohistology and gene-expression analysis was carried out on freshly isolated cells and cultured cells. Cell morphology and proliferation were analysed by two-photon-laser-microscopy. Two-photon-laser-microscopy showed a homogenous and small CLC population in the inner annulus, which differed from the large vacuole-containing NC in the nucleus. Immunohistology found 93 % KRT8 positive cells in the nucleus and intracellular and pericellular Col2, IL6, and IL12 staining while CLC were KRT8 negative. Freshly isolated NC showed significantly higher KRT8 and CAIII but lower Col2 gene-expression than CLC. NC in 3D-cultures demonstrated significant size reduction and loss of vacuoles with culture time, all indicating a loss of the characteristic NC morphology. Hypoxia reduced the rate of decrease in NC size and vacuoles. Gene-expression of KRT8 and CAIII in NC fell significantly early in culture while Col2 did not decrease significantly within the culture period. In CLC, KRT8 and CAIII gene-expression was low and did not change noticeably in culture, whereas Col2 expression fell with time in culture. 3D-culture
-
Radiosensitivity of primary cultured fish cells with different ploidy
International Nuclear Information System (INIS)
Mitani, Hiroshi; Egami, Nobuo; Kobayashi, Hiromu.
1986-01-01
The radiosensitivity of primary cultured goldfish cells (Carassius auratus) was investigated by colony formation assay. The radiosensitivity of cells from two varieties of goldfish, which show different sensitivity to lethal effect of ionizing radiation in vivo, was almost identical. Primary cultured cells from diploid, triploid and tetraploid fish retained their DNA content as measured by microfluorometry, and the nuclear size increases as ploidy increases. However, radiosensitivity was not related to ploidy. (author)
-
Neural differentiation of adipose-derived stem cells by indirect co-culture with Schwann cells
Directory of Open Access Journals (Sweden)
Li Xiaojie
2009-01-01
Full Text Available To investigate whether adipose-derived stem cells (ADSCs could be subject to neural differentiation induced only by Schwann cell (SC factors, we co-cultured ADSCs and SCs in transwell culture dishes. Immunoassaying, Western blot analysis, and RT-PCR were performed (1, 3, 7, 14 d and the co-cultured ADSCs showed gene and protein expression of S-100, Nestin, and GFAP. Further, qRT-PCR disclosed relative quantitative differences in the above three gene expressions. We think ADSCs can undergo induced neural differentiation by being co-cultured with SCs, and such differentiations begin 1 day after co-culture, become apparent after 7 days, and thereafter remain stable till the 14th day.
-
Microfluidic 3D cell culture: potential application for tissue-based bioassays
Li, XiuJun (James); Valadez, Alejandra V.; Zuo, Peng; Nie, Zhihong
2014-01-01
Current fundamental investigations of human biology and the development of therapeutic drugs, commonly rely on two-dimensional (2D) monolayer cell culture systems. However, 2D cell culture systems do not accurately recapitulate the structure, function, physiology of living tissues, as well as highly complex and dynamic three-dimensional (3D) environments in vivo. The microfluidic technology can provide micro-scale complex structures and well-controlled parameters to mimic the in vivo environment of cells. The combination of microfluidic technology with 3D cell culture offers great potential for in vivo-like tissue-based applications, such as the emerging organ-on-a-chip system. This article will review recent advances in microfluidic technology for 3D cell culture and their biological applications. PMID:22793034
-
The Evolution of Polystyrene as a Cell Culture Material.
Lerman, Max J; Lembong, Josephine; Muramoto, Shin; Gillen, Greg; Fisher, John P
2018-04-10
Polystyrene (PS) has brought in vitro cell culture from its humble beginnings to the modern era, propelling dozens of research fields along the way. This review discusses the development of the material, fabrication, and treatment approaches to create the culture material. However, native PS surfaces poorly facilitate cell adhesion and growthin vitro. To overcome this, liquid surface deposition, energetic plasma activation, and emerging functionalization methods transform the surface chemistry. This review seeks to highlight the many potential applications of the first widely accepted polymer growth surface. Although the majority of in vitro research occurs on 2D surfaces, the importance of 3D culture models cannot be overlooked. Here the methods to transition PS to specialized 3D culture surfaces are also reviewed. Specifically, casting, electrospinning, 3D printing, and microcarrier approaches to shift PS to a 3D culture surface are highlighted. The breadth of applications of the material makes it impossible to highlight every use, but the aim remains to demonstrate the versatility and potential as both a general and custom cell culture surface. The review concludes with emerging scaffolding approaches and, based on the findings, presents our insights on the future steps for PS as a tissue culture platform.
-
Cell death in Tetrahymena thermophila: new observations on culture conditions.
Christensen, S T; Sørensen, H; Beyer, N H; Kristiansen, K; Rasmussen, L; Rasmussen, M I
2001-01-01
We previously suggested that the cell fate of the protozoan ciliate, Tetrahymena thermophila, effectively relates to a quorum-sensing mechanism where cell-released factors support cell survival and proliferation. The cells have to be present above a critical initial density in a chemically defined nutrient medium in order to release a sufficient level of these factors to allow a new colony to flourish. At a relatively high rate of metabolism and/or macromolecular synthesis and below this critical density, cells began to die abruptly within 30 min of inoculation, and this death took the form of an explosive disintegration lasting less than 50 milliseconds. The cells died at any location in the culture, and the frequency of cell death was always lower in well-filled vials than those with medium/air interface. Cell death was inhibited by the addition of Actinomycin D or through modifications of the culture conditions either by reducing the oxygen tension or by decreasing the temperature of the growth medium. In addition, plastic caps in well-filled vials release substances, which promote cell survival. The fate of low-density cultures is related to certain 'physical' conditions, in addition to the availability of oxygen within closed culture systems. Copyright 2001 Academic Press.
-
A method for culturing human hair follicle cells.
Weterings, P J; Vermorken, A J; Bloemendal, H
1981-01-01
For the first time a method for culturing human hair follicle cells is described. The bovine eye lens capsule, a basement membrane-like structure, is used as the substrate for the cultures. In a culture medium supplemented with hydrocortisone and insulin about 70% of the original follicles will form growing colonies of diploid keratinocytes.
-
Improved Performance in Mammalian Cell Perfusion Cultures by Growth Inhibition.
Wolf, Moritz K F; Closet, Aurélie; Bzowska, Monika; Bielser, Jean-Marc; Souquet, Jonathan; Broly, Hervé; Morbidelli, Massimo
2018-05-21
Mammalian cell perfusion cultures represent a promising alternative to the current fed-batch technology for the production of various biopharmaceuticals. Long-term operation at a fixed viable cell density (VCD) requires a viable culture and a constant removal of excessive cells. Product loss in the cell removing bleed stream deteriorates the process yield. In this study, the authors investigate the use of chemical and environmental growth inhibition on culture performance by either adding valeric acid (VA) to the production media or by reducing the culture temperature (33.0 °C) with respect to control conditions (36.5 °C, no VA). Low temperature significantly reduces cellular growth, thus, resulting in lower bleed rates accompanied by a reduced product loss of 11% compared to 26% under control conditions. Additionally, the cell specific productivity of the target protein improves and maintained stable leading to media savings per mass of product. VA shows initially an inhibitory effect on cellular growth. However, cells seemed to adapt to the presence of the inhibitor resulting in a recovery of the cellular growth. Cell cycle and Western blot analyses support the observed results. This work underlines the role of temperature as a key operating variable for the optimization of perfusion cultures. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
-
Self-assembling Fmoc dipeptide hydrogel for in situ 3D cell culturing
Directory of Open Access Journals (Sweden)
Akpe Victor
2007-12-01
Full Text Available Abstract Background Conventional cell culture studies have been performed on 2D surfaces, resulting in flat, extended cell growth. More relevant studies are desired to better mimic 3D in vivo tissue growth. Such realistic environments should be the aim of any cell growth study, requiring new methods for culturing cells in vitro. Cell biology is also tending toward miniaturization for increased efficiency and specificity. This paper discusses the application of a self-assembling peptide-derived hydrogel for use as a 3D cell culture scaffold at the microscale. Results Phenylalanine derivative hydrogel formation was seen to occur in multiple dispersion media. Cells were immobilized in situ within microchambers designed for cell analysis. Use of the highly biocompatible hydrogel components and simplistic procedures significantly reduced the cytotoxic effects seen with alternate 3D culture materials and microstructure loading methods. Cells were easily immobilized, sustained and removed from microchambers. Differences in growth morphology were seen in the cultured cells, owing to the 3-dimentional character of the gel structure. Degradation improved the removal of hydrogel from the microstructures, permitting reuse of the analysis platforms. Conclusion Self-assembling diphenylalanine derivative hydrogel provided a method to dramatically reduce the typical difficulties of microculture formation. Effective generation of patterned 3D cultures will lead to improved cell study results by better modeling in vivo growth environments and increasing efficiency and specificity of cell studies. Use of simplified growth scaffolds such as peptide-derived hydrogel should be seen as highly advantageous and will likely become more commonplace in cell culture methodology.
-
Krawczyk, Krzysztof M; Matak, Damian; Szymanski, Lukasz; Szczylik, Cezary; Porta, Camillo; Czarnecka, Anna M
2018-04-01
The use of fetal bovine serum hinders obtaining reproducible experimental results and should also be removed in hormone and growth factor studies. In particular hormones found in FBS act globally on cancer cell physiology and influence transcriptome and metabolome. The aim of our study was to develop a renal carcinoma serum free culture model optimized for (embryonal) renal cells in order to select the best study model for downstream auto-, para- or endocrine research. Secondary aim was to verify renal carcinoma stem cell culture for this application. In the study, we have cultured renal cell carcinoma primary tumour cell line (786-0) as well as human kidney cancer stem cells in standard 2D monolayer cultures in Roswell Park Memorial Institute Medium or Dulbecco's Modified Eagle's Medium and Complete Human Kidney Cancer Stem Cell Medium, respectively. Serum-free, animal-component free Human Embryonic Kidney 293 media were tested. Our results revealed that xeno-free embryonal renal cells optimized culture media provide a useful tool in RCC cancer biology research and at the same time enable effective growth of RCC. We propose bio-mimic RCC cell culture model with specific serum-free and xeno-free medium that promote RCC cell viability.
-
Sensor Access to the Cellular Microenvironment Using the Sensing Cell Culture Flask.
Kieninger, Jochen; Tamari, Yaara; Enderle, Barbara; Jobst, Gerhard; Sandvik, Joe A; Pettersen, Erik O; Urban, Gerald A
2018-04-26
The Sensing Cell Culture Flask (SCCF) is a cell culture monitoring system accessing the cellular microenvironment in 2D cell culture using electrochemical microsensors. The system is based on microfabricated sensor chips embedded in standard cell culture flasks. Ideally, the sensor chips could be equipped with any electrochemical sensor. Its transparency allows optical inspection of the cells during measurement. The surface of the sensor chip is in-plane with the flask surface allowing undisturbed cell growth on the sensor chip. A custom developed rack system allows easy usage of multiple flasks in parallel within an incubator. The presented data demonstrates the application of the SCCF with brain tumor (T98G) and breast cancer (T-47D) cells. Amperometric oxygen sensors were used to monitor cellular respiration with different incubation conditions. Cellular acidification was accessed with potentiometric pH sensors using electrodeposited iridium oxide films. The system itself provides the foundation for electrochemical monitoring systems in 3D cell culture.
-
Sensor Access to the Cellular Microenvironment Using the Sensing Cell Culture Flask
Directory of Open Access Journals (Sweden)
Jochen Kieninger
2018-04-01
Full Text Available The Sensing Cell Culture Flask (SCCF is a cell culture monitoring system accessing the cellular microenvironment in 2D cell culture using electrochemical microsensors. The system is based on microfabricated sensor chips embedded in standard cell culture flasks. Ideally, the sensor chips could be equipped with any electrochemical sensor. Its transparency allows optical inspection of the cells during measurement. The surface of the sensor chip is in-plane with the flask surface allowing undisturbed cell growth on the sensor chip. A custom developed rack system allows easy usage of multiple flasks in parallel within an incubator. The presented data demonstrates the application of the SCCF with brain tumor (T98G and breast cancer (T-47D cells. Amperometric oxygen sensors were used to monitor cellular respiration with different incubation conditions. Cellular acidification was accessed with potentiometric pH sensors using electrodeposited iridium oxide films. The system itself provides the foundation for electrochemical monitoring systems in 3D cell culture.
-
Butler, W B
1984-08-15
A procedure is described for preparing nuclei from cells in monolayer culture so that they may be counted using an electronic particle counter. It takes only 10 to 15 min, and consists of swelling the cells in hypotonic buffer and then lysing them with the quaternary ammonium salt, ethylhexadecyldimethylammonium bromide. The cells are completely lysed, yielding a suspension of clean single nuclei which is stable, free of debris, and easily counted. The method was developed for a cell line of epithelial origin (MCF-7), which is often difficult to trypsinize to single cells. It works equally well at all cell densities up to and beyond confluence, and has been used with a variety of cells in culture, including 3T3 cells, bovine macrophages, rat mammary epithelial cells, mouse mammary tumor cell lines, and human fibroblasts. The size of the nuclei produced by this procedure is related to their DNA content, and the method is thus suitable for following cultures of synchronized cells through the cell cycle, and for performing differential counts of cells with substantial differences in DNA content.
-
MAPLE deposition of 3D micropatterned polymeric substrates for cell culture
Energy Technology Data Exchange (ETDEWEB)
Paun, Irina Alexandra, E-mail: irina.paun@physics.pub.ro [National Institute for Laser, Plasma and Radiation Physics, RO-077125, Magurele, Bucharest (Romania); Faculty of Applied Sciences, University Politehnica of Bucharest, RO-060042, Bucharest (Romania); Mihailescu, Mona [Faculty of Applied Sciences, University Politehnica of Bucharest, RO-060042, Bucharest (Romania); Calenic, Bogdan [Department of Biochemistry, Faculty of Dentistry, UMF Carol Davila, Bucharest (Romania); Luculescu, Catalin Romeo [National Institute for Laser, Plasma and Radiation Physics, RO-077125, Magurele, Bucharest (Romania); Greabu, Maria [Department of Biochemistry, Faculty of Dentistry, UMF Carol Davila, Bucharest (Romania); Dinescu, Maria, E-mail: dinescum@nipne.ro [National Institute for Laser, Plasma and Radiation Physics, RO-077125, Magurele, Bucharest (Romania)
2013-08-01
3D micropatterned poly(lactide-co-glycolide)/polyurethane (PLGA/PU) substrates were produced by MAPLE deposition through masks and used for regulating the behavior of oral keratinocyte stem cells in response to topography. Flat PLGA/PU substrates were produced for comparison. 3D imaging of the PLGA/PU substrates and of the cultured cells was performed by Digital Holographic Microscopy. The micropatterns were in the shape of squares of 50 × 50 and 80 × 80 μm{sup 2} areas, ∼1.8 μm in height and separated by 20 μm wide channels. It was found that substrate topography guided the adhesion of the cultured cells: on the smooth substrates the cells adhered randomly and showed no preferred orientation; in contrast, on the micropatterned substrates the cells adhered preferentially onto the squares and not in the separating channels. Furthermore, key properties of the cells (size, viability, proliferation rate and stem cell marker expression) did not show any dependence on substrate topography. The size of the cultured cells, their viability, the proportions of actively/slow proliferating cells, as well as the stem cell markers expressions, were similar for both flat and micropatterned substrates. Finally, it was found that the cells cultured on the PLGA/PU substrates deposited by MAPLE exhibited similar properties as the controls (i.e. cells cultured on glass slides), indicating the capability of the former to preserve the properties of the keratinocyte stem cells.
-
Lingual Epithelial Stem Cells and Organoid Culture of Them
Directory of Open Access Journals (Sweden)
Hiroko Hisha
2016-01-01
Full Text Available As tongue cancer is one of the major malignant cancers in the world, understanding the mechanism of maintenance of lingual epithelial tissue, which is known to be the origin of tongue cancer, is unquestionably important. However, the actual stem cells that are responsible for the long-term maintenance of the lingual epithelium have not been identified. Moreover, a simple and convenient culture method for lingual epithelial stem cells has not yet been established. Recently, we have shown that Bmi1-positive cells, residing at the second or third layer of the epithelial cell layer at the base of the interpapillary pit (IPP, were slow-cycling and could supply keratinized epithelial cells for over one year, indicating that Bmi1-positive cells are long-term lingual epithelial stem cells. In addition, we have developed a novel lingual epithelium organoid culture system using a three-dimensional matrix and growth factors. Here, we discuss current progress in the identification of lingual stem cells and future applications of the lingual culture system for studying the regulatory mechanisms of the lingual epithelium and for regenerative medicine.
-
Between Faith and Science: World Culture Theory and Comparative Education
Carney, Stephen; Rappleye, Jeremy; Silova, Iveta
2012-01-01
World culture theory seeks to explain an apparent convergence of education through a neoinstitutionalist lens, seeing global rationalization in education as driven by the logic of science and the myth of progress. While critics have challenged these assumptions by focusing on local manifestations of world-level tendencies, such critique is…
-
Plant Cell Cultures as Source of Cosmetic Active Ingredients
Directory of Open Access Journals (Sweden)
Ani Barbulova
2014-04-01
Full Text Available The last decades witnessed a great demand of natural remedies. As a result, medicinal plants have been increasingly cultivated on a commercial scale, but the yield, the productive quality and the safety have not always been satisfactory. Plant cell cultures provide useful alternatives for the production of active ingredients for biomedical and cosmetic uses, since they represent standardized, contaminant-free and biosustainable systems, which allow the production of desired compounds on an industrial scale. Moreover, thanks to their totipotency, plant cells grown as liquid suspension cultures can be used as “biofactories” for the production of commercially interesting secondary metabolites, which are in many cases synthesized in low amounts in plant tissues and differentially distributed in the plant organs, such as roots, leaves, flowers or fruits. Although it is very widespread in the pharmaceutical industry, plant cell culture technology is not yet very common in the cosmetic field. The aim of the present review is to focus on the successful research accomplishments in the development of plant cell cultures for the production of active ingredients for cosmetic applications.
-
Establishment of an immortalized mouse dermal papilla cell strain with optimized culture strategy
Directory of Open Access Journals (Sweden)
Haiying Guo
2018-01-01
Full Text Available Dermal papilla (DP plays important roles in hair follicle regeneration. Long-term culture of mouse DP cells can provide enough cells for research and application of DP cells. We optimized the culture strategy for DP cells from three dimensions: stepwise dissection, collagen I coating, and optimized culture medium. Based on the optimized culture strategy, we immortalized primary DP cells with SV40 large T antigen, and established several immortalized DP cell strains. By comparing molecular expression and morphologic characteristics with primary DP cells, we found one cell strain named iDP6 was similar with primary DP cells. Further identifications illustrate that iDP6 expresses FGF7 and α-SMA, and has activity of alkaline phosphatase. During the process of characterization of immortalized DP cell strains, we also found that cells in DP were heterogeneous. We successfully optimized culture strategy for DP cells, and established an immortalized DP cell strain suitable for research and application of DP cells.
-
Hamuro, Junji; Toda, Munetoyo; Asada, Kazuko; Hiraga, Asako; Schlötzer-Schrehardt, Ursula; Montoya, Monty; Sotozono, Chie; Ueno, Morio; Kinoshita, Shigeru
2016-09-01
To identify the subpopulation (SP) among heterogeneous cultured human corneal endothelial cells (cHCECs) devoid of cell-state transition applicable for cell-based therapy. Subpopulation presence in cHCECs was confirmed via surface CD-marker expression level by flow cytometry. CD markers effective for distinguishing distinct SPs were selected by analyzing those on established cHCECs with a small cell area and high cell density. Contrasting features among three typical cHCEC SPs was confirmed by PCR array for extracellular matrix (ECM). Combined analysis of CD markers was performed to identify the SP (effector cells) applicable for therapy. ZO-1 and Na+/K+ ATPase, CD200, and HLA expression were compared among heterogeneous SPs. Flow cytometry analysis identified the effector cell expressing CD166+CD105-CD44-∼+/-CD26-CD24-, but CD200-, and the presence of other SPs with CD166+ CD105-CD44+++ (CD26 and CD24, either + or -) was confirmed. PCR array revealed three distinct ECM expression profiles. Some SPs expressed ZO-1 and Na+/K+ ATPase at comparable levels with effector cells, while only one SP expressed CD200, but not on effector cells. Human leukocyte antigen expression was most reduced in the effector SP. The proportion of effector cells (E-ratio) inversely paralleled donor age and decreased during prolonged culture passages. The presence of Rho-associated protein kinase (ROCK) inhibitor increased the E-ratio in cHCECs. The average area of effector cells was approximately 200∼220 μm2, and the density of cHCECs exceeded 2500 cells/mm2. A specified cultured effector cell population sharing the surface phenotypes with mature HCECs in corneal tissues may serve as an alternative to donor corneas for the treatment of corneal endothelial dysfunction.
-
Stevens, Sally; Andrade, Rosi; Page, Melissa
2016-01-01
Data indicate that females and ethnic/race minority groups are underrepresented in the science and engineering workforce calling for innovative strategies to engage and retain them in science education and careers. This study reports on the development, delivery, and outcomes of a culturally driven science, technology, engineering, mathematics…
-
Mouse cell culture - Methods and protocols
Directory of Open Access Journals (Sweden)
CarloAlberto Redi
2010-12-01
Full Text Available The mouse is, out of any doubt, the experimental animal par excellence for many many colleagues within the scientific community, notably for those working in mammalian biology (in a broad sense, from basic genetic to modeling human diseases, starting at least from 1664 Robert Hooke experiments on air’s propertyn. Not surprising then that mouse cell cultures is a well established field of research itself and that there are several handbooks devoted to this discipline. Here, Andrew Ward and David Tosh provide a necessary update of the protocols currently needed. In fact, nearly half of the book is devoted to stem cells culture protocols, mainly embryonic, from a list of several organs (kidney, lung, oesophagus and intestine, pancreas and liver to mention some........
-
Yoshida, Ryu; Cheng, Mingyu; Murray, Martha M
2014-02-01
Tissue engineering is one new strategy being developed to treat ACL ruptures. One such approach is bio-enhanced ACL repair, where a suture repair is supplemented with a bio-active scaffold containing platelets. However, the optimal concentration of platelets to stimulate ACL healing is not known. We hypothesized that increasing platelet concentrations in the scaffold would enhance critical cell behaviors. Porcine ACL fibroblasts were obtained from explant culture and suspended in platelet poor plasma (PPP), 1× platelet-rich plasma (PRP), 3× PRP, 5× PRP, or phosphate buffered saline (PBS). The cell suspensions were cultured in a 3D collagen scaffold. Cellular metabolism (MTT assay), apoptosis (TUNEL assay), and gene expression for type I and type III collagen were measured. 1× PRP significantly outperformed 5× PRP in all parameters studied: Type I and III collagen gene expression, apoptosis prevention, and cell metabolism stimulation. ACL fibroblasts cultured with 1× PRP had the highest type I and type III collagen gene expression. 1× PRP and PPP groups had the highest cell metabolism and lowest apoptosis rates. Concentration of platelets had significant effects on the behavior of ACL fibroblasts; thus, it is an important parameter that should be specified in clinical or basic science studies. © 2013 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.
-
Gehre, Nadine; Nusser, Anja; von Muenchow, Lilly; Tussiwand, Roxane; Engdahl, Corinne; Capoferri, Giuseppina; Bosco, Nabil; Ceredig, Rhodri; Rolink, Antonius G
2015-03-01
T-cell lymphopenia following BM transplantation or diseases such as AIDS result in immunodeficiency. Novel approaches to ameliorate this situation are urgently required. Herein, we describe a novel stromal cell free culture system in which Lineage(-) Sca1(+)c-kit(+) BM hematopoietic progenitors very efficiently differentiate into pro-T cells. This culture system consists of plate-bound Delta-like 4 Notch ligand and the cytokines SCF and IL-7. The pro-T cells developing in these cultures express CD25, CD117, and partially CD44; express cytoplasmic CD3ε; and have their TCRβ locus partially D-J rearranged. They could be expanded for over 3 months and used to reconstitute the T-cell compartments of sublethally irradiated T-cell-deficient CD3ε(-/-) mice or lethally irradiated WT mice. Pro-T cells generated in this system could partially correct the T-cell lymphopenia of pre-Tα(-/-) mice. However, reconstituted CD3ε(-/-) mice suffered from a wasting disease that was prevented by co-injection of purified CD4(+) CD25(high) WT Treg cells. In a T-cell-sufficient or T-lymphopenic setting, the development of disease was not observed. Thus, this in vitro culture system represents a powerful tool to generate large numbers of pro-T cells for transplantation and possibly with clinical applications. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
-
Energy Technology Data Exchange (ETDEWEB)
Stio, Maria; Martinesi, Maria; Treves, Cristina [Dipartimento di Scienze Biomediche, Sperimentali e Cliniche ‘Mario Serio’, Sezione di Scienze Biochimiche, Università di Firenze, viale Morgagni 50, 50134 Firenze (Italy); Borgioli, Francesca, E-mail: francesca.borgioli@unifi.it [Dipartimento di Ingegneria Industriale (DIEF), Università di Firenze, via S. Marta 3, 50139 Firenze (Italy)
2016-12-01
Samples of AISI 316L austenitic stainless steel were subjected either to grinding and polishing procedure, or to grinding and then low temperature glow-discharge nitriding treatment, or to grinding, nitriding and subsequently coating with collagen-I. Nitrided samples, even if only ground, show a higher corrosion resistance in PBS solution, in comparison with ground and polished AISI 316L. Biocompatibility was evaluated in vitro by incubating the samples with either peripheral blood mononuclear cells (PBMC) or human umbilical vein endothelial cells (HUVEC), tested separately or in co-culture. HUVEC-PBMC co-culture and co-incubation of HUVEC with PBMC culture medium, after the previous incubation of PBMC with metallic samples, allowed to determine whether the incubation of PBMC with the different samples might affect HUVEC behaviour. Many biological parameters were considered: cell proliferation, release of cytokines, matrix metalloproteinases (MMPs) and sICAM-1, gelatinolytic activity of MMPs, and ICAM-1 protein expression. Nitriding treatment, with or without collagen coating of the samples, is able to ameliorate some of the biological parameters taken into account. The obtained results point out that biocompatibility may be successfully tested in vitro, using cultures of normal human cells, as blood and endothelial cells, but more than one cell line should be used, separately or in co-culture, and different parameters should be determined, in particular those correlated with inflammatory phenomena. - Highlights: • Nitriding improves corrosion resistance and biocompatibility of ground AISI 316L. • The metallic samples differently affect different human cell cultures. • PBMC and HUVEC are a suitable model to test in vitro biocompatibility. • Co-cultures show that HUVEC are affected by pre-incubation of PBMC with the samples. • Inflammation parameters must be taken into account for assessing biocompatibility.
-
International Nuclear Information System (INIS)
Stio, Maria; Martinesi, Maria; Treves, Cristina; Borgioli, Francesca
2016-01-01
Samples of AISI 316L austenitic stainless steel were subjected either to grinding and polishing procedure, or to grinding and then low temperature glow-discharge nitriding treatment, or to grinding, nitriding and subsequently coating with collagen-I. Nitrided samples, even if only ground, show a higher corrosion resistance in PBS solution, in comparison with ground and polished AISI 316L. Biocompatibility was evaluated in vitro by incubating the samples with either peripheral blood mononuclear cells (PBMC) or human umbilical vein endothelial cells (HUVEC), tested separately or in co-culture. HUVEC-PBMC co-culture and co-incubation of HUVEC with PBMC culture medium, after the previous incubation of PBMC with metallic samples, allowed to determine whether the incubation of PBMC with the different samples might affect HUVEC behaviour. Many biological parameters were considered: cell proliferation, release of cytokines, matrix metalloproteinases (MMPs) and sICAM-1, gelatinolytic activity of MMPs, and ICAM-1 protein expression. Nitriding treatment, with or without collagen coating of the samples, is able to ameliorate some of the biological parameters taken into account. The obtained results point out that biocompatibility may be successfully tested in vitro, using cultures of normal human cells, as blood and endothelial cells, but more than one cell line should be used, separately or in co-culture, and different parameters should be determined, in particular those correlated with inflammatory phenomena. - Highlights: • Nitriding improves corrosion resistance and biocompatibility of ground AISI 316L. • The metallic samples differently affect different human cell cultures. • PBMC and HUVEC are a suitable model to test in vitro biocompatibility. • Co-cultures show that HUVEC are affected by pre-incubation of PBMC with the samples. • Inflammation parameters must be taken into account for assessing biocompatibility.
-
Skottman, H; Muranen, J; Lähdekorpi, H; Pajula, E; Mäkelä, K; Koivusalo, L; Koistinen, A; Uusitalo, H; Kaarniranta, K; Juuti-Uusitalo, K
2017-10-01
Here we evaluated the effects of human retinal microvascular endothelial cells (hREC) on mature human embryonic stem cell (hESC) derived retinal pigment epithelial (RPE) cells. The hESC-RPE cells (Regea08/017, Regea08/023 or Regea11/013) and hREC (ACBRI 181) were co-cultured on opposite sides of transparent membranes for up to six weeks. Thereafter barrier function, small molecule permeability, localization of RPE and endothelial cell marker proteins, cellular fine structure, and growth factor secretion of were evaluated. After co-culture, the RPE specific CRALBP and endothelial cell specific von Willebrand factor were appropriately localized. In addition, the general morphology, pigmentation, and fine structure of hESC-RPE cells were unaffected. Co-culture increased the barrier function of hESC-RPE cells, detected both with TEER measurements and cumulative permeability of FD4 - although the differences varied among the cell lines. Co-culturing significantly altered VEGF and PEDF secretion, but again the differences were cell line specific. The results of this study showed that co-culture with hREC affects hESC-RPE functionality. In addition, co-culture revealed drastic cell line specific differences, most notably in growth factor secretion. This model has the potential to be used as an in vitro outer blood-retinal barrier model for drug permeability testing. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.
-
International Nuclear Information System (INIS)
Ruemke, H.C.; Terpstra, F.G.; Huis, B.; Out, T.A.; Zeijlemaker, W.P.
1982-01-01
When human peripheral blood lymphocytes (PBL) are cultured in the presence of irradiated allogeneic lymphocytes, the resulting mixed lymphocyte reaction (MLR) leads to the secretion into the supernatant of substantial amounts of IgM and IgG, derived from nonirradiated responder B lymphocytes. Our data indicate that stimulation to Ig production by responder B cells may result from different types of of interactions. First, B cells and monocytes among the irradiated stimulator cells activate T responder B cells to produce Ig; second, ''responder'' B cells activate irradiated ''stimulator'' T cells, leading to a ''helper'' signal, back to the responder B cells and leading to Ig production. The latter system is radiosensitive, because allogeneic T cells, irradiated at a dose of 4000 rad or more, failed to induce Ig production by responder B cells. In some combinations of human allogeneic lymphocytes, the co-culture of the cells leads to inhibition of Ig production, both in the presence and in the absence of PWM. Thus, co-culture of allogeneic cells may cause ''positive'' as well as ''negative'' allogeneic effects. The implications of these findings for the interpretation of co-cultures that are aimed at establishing defects in lymphocytes from patients with, for example, immunodeficiencies, who fail to produce Ig in the presence of PWM are discussed
-
Control of fibronectin synthesis by rat granulosa cells in culture
International Nuclear Information System (INIS)
Skinner, M.K.; Dorrington, J.H.
1984-01-01
The secreted and cellular [ 35 S]methionine-radiolabeled proteins of cultured rat granulosa cells were separated by electrophoresis on sodium dodecylsulfate (SDS) polyacrylamide gradient slab gels. From 24 to 72 h of culture FSH increased the intensity of labeling of most of the secreted proteins. A 220,000-dalton protein, however, increased in intensity only in control cultures and became the major secreted protein after 72 h, comprising 20% of the total radiolabeled proteins. This protein was identified as fibronectin by immunoprecipitation. There was no increase in the secreted or cellular fibronectin in FSH- or testosterone- and insulin-treated cultures. These studies indicate that a component of extracellular matrix is a major secretory product of unstimulated immature granulosa cells. As hormones induce the differentiated functions of granulosa cells in culture, the secretion of fibronectin is inhibited
-
Differential heat shock response of primary human cell cultures and established cell lines
DEFF Research Database (Denmark)
Richter, W W; Issinger, O G
1986-01-01
degrees C treatment, whereas in immortalized cell lines usually 90% of the cells were found in suspension. Enhanced expression of the major heat shock protein (hsp 70) was found in all heat-treated cells. In contrast to the primary cell cultures, established and transformed cell lines synthesized...
-
Berry, Joseph D; Godara, Pankaj; Liovic, Petar; Haylock, David N
2015-04-16
Recent studies in the literature have highlighted the critical role played by cell signalling in determining haemopoietic stem cell (HSC) fate within ex vivo culture systems. Stimulatory signals can enhance proliferation and promote differentiation, whilst inhibitory signals can significantly limit culture output. Numerical models of various mitigation strategies are presented and applied to determine effectiveness of these strategies toward mitigation of paracrine inhibitory signalling inherent in these culture systems. The strategies assessed include mixing, media-exchange, fed-batch and perfusion. The models predict that significant spatial concentration gradients exist in typical cell cultures, with important consequences for subsequent cell expansion. Media exchange is shown to be the most effective mitigation strategy, but remains labour intensive and difficult to scale-up for large culture systems. The fed-batch strategy is only effective at very small Peclet number, and its effect is diminished as the cell culture volume grows. Conversely, mixing is effective at high Peclet number, and ineffective at low Peclet number. The models predict that cell expansion in fed-batch cultures becomes independent of increasing dilution rate, consistent with experimental results previously reported in the literature. In contrast, the models predict that increasing the flow rate in perfused cultures will lead to increased cell expansion, indicating the suitability of perfusion for use as an automated, tunable strategy. The effect of initial cell seeding density is also investigated, with the model showing that perfusion outperforms dilution for all densities considered. The models predict that the impact of inhibitory signalling in HSC cultures can be mitigated against using media manipulation strategies, with the optimal strategy dependent upon the protein diffusion time-scale relative to the media manipulation time-scale. The key messages from this study can be applied to
-
Human disc cells in monolayer vs 3D culture: cell shape, division and matrix formation
Directory of Open Access Journals (Sweden)
Hanley Edward N
2000-10-01
Full Text Available Abstract Background The relationship between cell shape, proliferation, and extracellular matrix (ECM production, important aspects of cell behavior, is examined in a little-studied cell type, the human annulus cell from the intervertebral disc, during monolayer vs three-dimensional (3D culture. Results Three experimental studies showed that cells respond specifically to culture microenvironments by changes in cell shape, mitosis and ECM production: 1 Cell passages showed extensive immunohistochemical evidence of Type I and II collagens only in 3D culture. Chondroitin sulfate and keratan sulfate were abundant in both monolayer and 3D cultures. 2 Cells showed significantly greater proliferation in monolayer in the presence of platelet-derived growth factor compared to cells in 3D. 3 Cells on Matrigel™-coated monolayer substrates became rounded and formed nodular colonies, a finding absent during monolayer growth. Conclusions The cell's in vivo interactions with the ECM can regulate shape, gene expression and other cell functions. The shape of the annulus cell changes markedly during life: the young, healthy disc contains spindle shaped cells and abundant collagen. With aging and degeneration, many cells assume a strikingly different appearance, become rounded and are surrounded by unusual accumulations of ECM products. In vitro manipulation of disc cells provides an experimental window for testing how disc cells from given individuals respond when they are grown in environments which direct cells to have either spindle- or rounded-shapes. In vitro assessment of the response of such cells to platelet-derived growth factor and to Matrigel™ showed a continued influence of cell shape even in the presence of a growth factor stimulus. These findings contribute new information to the important issue of the influence of cell shape on cell behavior.
-
Paula M. Pijut; R. Daniel Lineberger; Subhash C. Domir; Jann M. Ichida; Charles R. Krause
1990-01-01
Calli of American elm susceptible and resistant to Dutch elm disease were exposed to a culture filtrate of a pathogenic isolate of Ceratocystis ulmi. Cells from untreated tissue exhibited typical internal composition associated with healthy, actively growing cells. All cells exposed to culture filtrate showed appreciable ultrastructural changes....
-
Marxism in Vygotskian approaches to cultural studies of science education
Lima Junior, Paulo; Ostermann, Fernanda; Rezende, Flavia
2014-09-01
In this paper we initially address the main categories of Marxism, illustrating how Vygotsky has appropriated them as mediational meta-theoretical tools for building concepts for his psychological approach. In order to investigate the influence of Marxism in cultural studies of science education, we make an account of how current research, sustained by Vygotsky's original and successor theories, has been appropriating meta-theoretical categories of dialectical materialism. Once we identified Cultural Studies of Science Education as a journal that would probably concentrate papers that follow these perspectives, we decided to take it as the context of this study. In the process of selecting the corpus to be reviewed from the editions published from 2006 to 2011, we have found that 16 % of the articles that matched keywords denoting frameworks related to the Vygotskian tradition developed and appropriated the categories of dialectical materialism. The quality and originality of contemporary development of CHAT denote that this framework has been playing a very important role in recent expansion of Vygotskian approaches to research in science education. Among the papers that we considered to develop and appropriate Vygotskian frameworks, incompletion in the appropriation of meta-theoretical categories of dialectical materialism and the misusage of dialectics intertwined with dialogism were highlighted. Our findings suggest that overcoming these limitations can enhance political analysis of sociocultural phenomena in the context of science education. It also represents a strengthening of the role of dialectical materialism in expanding sociocultural perspectives toward a better articulation between individual and institutional-centered analyses.
-
International Nuclear Information System (INIS)
Katano, Takahito; Ootani, Akifumi; Mizoshita, Tsutomu; Tanida, Satoshi; Tsukamoto, Hironobu; Ozeki, Keiji; Ebi, Masahide; Mori, Yoshinori; Kataoka, Hiromi; Kamiya, Takeshi; Toda, Shuji; Joh, Takashi
2013-01-01
Highlights: ► We established a 3D culture system to allow long-term culture of stomach cells. ► In this culture system, gastric epithelial cells grew for about 3 months. ► The cultured cells differentiated into multi-units of the stomach. ► This culture method should be useful for elucidating the cause of gastric diseases. -- Abstract: Compared to the small intestine and colon, little is known about stem cells in the stomach because of a lack of specific stem cell markers and an in vitro system that allows long-term culture. Here we describe a long-term three-dimensional (3D) primary gastric culture system within the stem cell niche. Glandular stomach cells from neonatal mice cultured in collagen gel yielded expanding sphere-like structures for 3 months. The wall of the gastrospheres consisted of a highly polarized epithelial monolayer with an outer lining of myofibroblasts. The epithelial cells showed a tall columnar cell shape, basal round nuclei, and mucus-filled cytoplasm as well as expression of MUC5AC, indicating differentiation into gastric surface mucous cells. These cells demonstrated the features of fully differentiated gastric surface mucous cells such as microvilli, junctional complexes, and glycogen and secretory granules. Fewer than 1% of cultured epithelial cells differentiated into enteroendocrine cells. Active proliferation of the epithelial cells and many apoptotic cells in the inner lumen revealed the rapid cell turnover in gastrospheres in vitro. This method enables us to investigate the role of signaling between cell–cell and epithelial–mesenchymal interactions in an environment that is extremely similar to the in vivo environment
-
Energy Technology Data Exchange (ETDEWEB)
Katano, Takahito [Department of Gastroenterology and Metabolism, Nagoya City University Graduate School of Medical Sciences, 1 Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya 467-8601 (Japan); Ootani, Akifumi [Department of Gastroenterology and GI Endoscopy Center, Shin-Kokura Hospital, Federation of National Public Service Personnel Mutual Aid Associations, 1-3-1 Kanada, Kokurakita-ku, Kitakyushu 803-0816 (Japan); Department of Internal Medicine, Faculty of Medicine, Saga University, 5-1-1 Nabeshima, Saga 849-8501 (Japan); Mizoshita, Tsutomu, E-mail: tmizoshi@med.nagoya-cu.ac.jp [Department of Gastroenterology and Metabolism, Nagoya City University Graduate School of Medical Sciences, 1 Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya 467-8601 (Japan); Tanida, Satoshi; Tsukamoto, Hironobu; Ozeki, Keiji; Ebi, Masahide; Mori, Yoshinori; Kataoka, Hiromi; Kamiya, Takeshi [Department of Gastroenterology and Metabolism, Nagoya City University Graduate School of Medical Sciences, 1 Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya 467-8601 (Japan); Toda, Shuji [Department of Pathology and Microbiology, Faculty of Medicine, Saga University, 5-1-1 Nabeshima, Saga 849-8501 (Japan); Joh, Takashi [Department of Gastroenterology and Metabolism, Nagoya City University Graduate School of Medical Sciences, 1 Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya 467-8601 (Japan)
2013-03-22
Highlights: ► We established a 3D culture system to allow long-term culture of stomach cells. ► In this culture system, gastric epithelial cells grew for about 3 months. ► The cultured cells differentiated into multi-units of the stomach. ► This culture method should be useful for elucidating the cause of gastric diseases. -- Abstract: Compared to the small intestine and colon, little is known about stem cells in the stomach because of a lack of specific stem cell markers and an in vitro system that allows long-term culture. Here we describe a long-term three-dimensional (3D) primary gastric culture system within the stem cell niche. Glandular stomach cells from neonatal mice cultured in collagen gel yielded expanding sphere-like structures for 3 months. The wall of the gastrospheres consisted of a highly polarized epithelial monolayer with an outer lining of myofibroblasts. The epithelial cells showed a tall columnar cell shape, basal round nuclei, and mucus-filled cytoplasm as well as expression of MUC5AC, indicating differentiation into gastric surface mucous cells. These cells demonstrated the features of fully differentiated gastric surface mucous cells such as microvilli, junctional complexes, and glycogen and secretory granules. Fewer than 1% of cultured epithelial cells differentiated into enteroendocrine cells. Active proliferation of the epithelial cells and many apoptotic cells in the inner lumen revealed the rapid cell turnover in gastrospheres in vitro. This method enables us to investigate the role of signaling between cell–cell and epithelial–mesenchymal interactions in an environment that is extremely similar to the in vivo environment.
-
Adherence of Moraxella bovis to cell cultures of bovine origin.
Annuar, B O; Wilcox, G E
1985-09-01
The adherence of five strains of Moraxella bovis to cell cultures was investigated. M bovis adhered to cultures of bovine corneal epithelial and Madin-Darby bovine kidney cells but not to cell types of non-bovine origin. Both piliated and unpiliated strains adhered but piliated strains adhered to a greater extent than unpiliated strains. Antiserum against pili of one strain inhibited adherence of piliated strains but caused only slight inhibition of adherence to the unpiliated strains. Treatment of bacteria with magnesium chloride caused detachment of pili from the bacterial cell and markedly inhibited adherence of piliated strains but caused only slight inhibition of adherence by the unpiliated strains. The results suggested that adhesion of piliated strains to cell cultures was mediated via pili but that adhesins other than pili may be involved in the attachment of unpiliated strains of M bovis to cells.
-
Science in Hawaii/Haawina Hoopapau: A Culturally Responsive Curriculum Project
Galloway, L. M.; Roberts, K.; Leake, D. W.; Stodden, R. S.; Crabbe, V.
2005-12-01
The marvels of modern science often fail to engage indigenous students, as the content and instructional style are usually rooted in the Western experience. This 3 year project, funded by the US Dept. of Education for the Education of Native Hawaiians, offers a curriculum that teaches science through (rather than just about) Native Hawaiian culture. The curriculum focuses on the interdependence of natural resources in our ahupuaa, or watersheds, and helps students strengthen their sense of place and self to malama i ka aina, to care for the land. Further, the curriculum is designed to: engage students in scientific study with relevant, interesting content and activities; improve student achievement of state department of education standards; increase student knowledge and skills in science, math and language arts; respond to the learning needs of Native Hawaiian and/or at-risk students. The project will be presented by a curriculum writer who created and adapted more than a year's worth of materials by teaming with kupuna (respected elders), local cultural experts and role models, educators (new, veteran, Hawaiian, non-Hawaiian, mainland, general and special education teachers), and professionals at the Center on Disability Studies at the University of Hawaii and ALU LIKE, Inc, a non-profit organization to assist Native Hawaiians. The materials created thus far are available for viewing at: www.scihi.hawaii.edu The curriculum, designed for grades 8-11 science classes, can be used to teach a year-long course, a unit, or single lesson related to astronomy, biology, botany, chemistry, geology, oceanography, physical and environmental sciences. This project is in its final year of field testing, polishing and dissemination, and therefore this session will encourage idea sharing, as does our copyright free Web site.
-
Quang, Tara; Marquez, Maribel; Blanco, Giselle; Zhao, Yuanxiang
2014-01-01
Many different culture systems have been developed for expanding human pluripotent stem cells (hESCs and hiPSCs). In general, 4-10 ng/ml of bFGF is supplemented in culture media in feeder-dependent systems regardless of feeder cell types, whereas in feeder-free systems, up to 100 ng/ml of bFGF is required for maintaining long-term culture on various substrates. The amount of bFGF required in native hESCs growth niche is unclear. Here we report using inactivated adipose-derived human mesenchymal stem cells as feeder cells to examine long-term parallel cultures of two hESCs lines (H1 and H9) and one hiPSCs line (DF19-9-7T) in media supplemented with 0, 0.4 or 4 ng/ml of bFGF for up to 23 passages, as well as parallel cultures of H9 and DF19 in media supplemented with 4, 20 or 100 ng/ml bFGF for up to 13 passages for comparison. Across all cell lines tested, bFGF supplement demonstrated inhibitory effect over growth expansion, single cell colonization and recovery from freezing in a dosage dependent manner. In addition, bFGF exerted differential effects on different cell lines, inducing H1 and DF19 differentiation at 4 ng/ml or higher, while permitting long-term culture of H9 at the same concentrations with no apparent dosage effect. Pluripotency was confirmed for all cell lines cultured in 0, 0.4 or 4 ng/ml bFGF excluding H1-4 ng, as well as H9 cultured in 4, 20 and 100 ng/ml bFGF. However, DF19 demonstrated similar karyotypic abnormality in both 0 and 4 ng/ml bFGF media while H1 and H9 were karyotypically normal in 0 ng/ml bFGF after long-term culture. Our results indicate that exogenous bFGF exerts dosage and cell line dependent effect on human pluripotent stem cells cultured on mesenchymal stem cells, and implies optimal use of bFGF in hESCs/hiPSCs culture should be based on specific cell line and its culture system.
-
Yi, Soyeon; Jang, Hyun-Jin; Lee, Hyo Suk; Yu, Jong-Phil; Kim, Soyeon; Lee, Joohee; Hur, Hee-Young
2013-06-01
In this study, we analyze the economic effects from the Korean Astronaut Program (KAP) and the subsequent Science Culture Diffusion Activity (SCDA). Korea has had a huge practical effect on the development of science and technology and has increased international awareness of Korea by producing Korea's first astronaut. There has also been a large, ripple effect on space related industries. In addition, the KAP has exercised a far-reaching influence on Korean society and culture by boosting all science and engineering and inspiring national pride. After the KAP, astronauts' outreach activities, such as lectures for the general public; interviews on television, newspapers and magazines; participating in children's science camps; and distributing publications and DVDs about astronaut program for general public, were instituted for diffusing science culture. Thus, positive effects such as the promotion of Korea's level of technology, student interest in science and engineering fields, and the expansion of the industrial base were reinforced after the KAP. This study is aimed at evaluating the economic significance and the value of return through analyzing the effects of the KAP and the subsequent Science Culture Diffusion Activity.
-
Marxism in Vygotskian Approaches to Cultural Studies of Science Education
Lima, Paulo, Jr.; Ostermann, Fernanda; Rezende, Flavia
2014-01-01
In this paper we initially address the main categories of Marxism, illustrating how Vygotsky has appropriated them as mediational meta-theoretical tools for building concepts for his psychological approach. In order to investigate the influence of Marxism in cultural studies of science education, we make an account of how current research,…
-
AMMONIA REMOVAL FROM MAMMALIAN CELL CULTURE MEDIUM BY ION-EXCHANGE MEMBRANES
Metabolites such as ammonia and lactic acid formed during mammalian cell culture can frequently be toxic to the cells themselves beyond a threshold concentration of the metabolites. Cell culture conducted in the presence of such accumulated metabolites is therefore limited in pro...
-
Advanced three-dimensional culture of equine intestinal epithelial stem cells.
Stewart, A Stieler; Freund, J M; Gonzalez, L M
2018-03-01
Intestinal epithelial stem cells are critical to epithelial repair following gastrointestinal injury. The culture of intestinal stem cells has quickly become a cornerstone of a vast number of new research endeavours that range from determining tissue viability to testing drug efficacy for humans. This study aims to describe the methods of equine stem cell culture and highlights the future benefits of these techniques for the advancement of equine medicine. To describe the isolation and culture of small intestinal stem cells into three-dimensional (3D) enteroids in horses without clinical gastrointestinal abnormalities. Descriptive study. Intestinal samples were collected by sharp dissection immediately after euthanasia. Intestinal crypts containing intestinal stem cells were dissociated from the underlying tissue layers, plated in a 3D matrix and supplemented with growth factors. After several days, resultant 3D enteroids were prepared for immunofluorescent imaging and polymerase chain reaction (PCR) analysis to detect and characterise specific cell types present. Intestinal crypts were cryopreserved immediately following collection and viability assessed. Intestinal crypts were successfully cultured and matured into 3D enteroids containing a lumen and budding structures. Immunofluorescence and PCR were used to confirm the existence of stem cells and all post mitotic, mature cell types, described to exist in the horse intestinal epithelium. Previously frozen crypts were successfully cultured following a freeze-thaw cycle. Tissues were all derived from normal horses. Application of this technique for the study of specific disease was not performed at this time. The successful culture of equine intestinal crypts into 3D "mini-guts" allows for in vitro studies of the equine intestine. Additionally, these results have relevance to future development of novel therapies that harness the regenerative potential of equine intestine in horses with gastrointestinal disease
-
Assessment of long-term effects of nanoparticles in a microcarrier cell culture system.
Directory of Open Access Journals (Sweden)
Maria Mrakovcic
Full Text Available Nano-sized materials could find multiple applications in medical diagnosis and therapy. One main concern is that engineered nanoparticles, similar to combustion-derived nanoparticles, may cause adverse effects on human health by accumulation of entire particles or their degradation products. Chronic cytotoxicity must therefore be evaluated. In order to perform chronic cytotoxicity testing of plain polystyrene nanoparticles on the endothelial cell line EAhy 926, we established a microcarrier cell culture system for anchorage-dependent cells (BioLevitator(TM. Cells were cultured for four weeks and exposed to doses, which were not cytotoxic upon 24 hours of exposure. For comparison, these particles were also studied in regularly sub-cultured cells, a method that has traditionally been used to assess chronic cellular effects. Culturing on basal membrane coated microcarriers produced very high cell densities. Fluorescent particles were mainly localized in the lysosomes of the exposed cells. After four weeks of exposure, the number of cells exposed to 20 nm polystyrene particles decreased by 60% as compared to untreated controls. When tested in sub-cultured cells, the same particles decreased cell numbers to 80% of the untreated controls. Dose-dependent decreases in cell numbers were also noted after exposure of microcarrier cultured cells to 50 nm short multi-walled carbon nanotubes. Our findings support that necrosis, but not apoptosis, contributed to cell death of the exposed cells in the microcarrier culture system. In conclusion, the established microcarrier model appears to be more sensitive for the identification of cellular effects upon prolonged and repeated exposure to nanoparticles than traditional sub-culturing.
-
Isolation, culture and adenoviral transduction of parietal cells from mouse gastric mucosa
International Nuclear Information System (INIS)
Gliddon, Briony L; Nguyen, Nhung V; Gunn, Priscilla A; Gleeson, Paul A; Driel, Ian R van
2008-01-01
Here we describe a method for the isolation of intact gastric glands from mice and primary culture and transfection of mouse gastric epithelial cells. Collagenase digestion of PBS-perfused mouse stomachs released large intact gastric glands that were plated on a basement membrane matrix. The heterogeneous gland cell cultures typically contain ∼60% parietal cells. Isolated mouse parietal cells remain viable in culture for up to 5 days and react strongly with an antibody specific to the gastric H + /K + ATPase. Isolated intact mouse gastric glands and primary cultures of mouse parietal cells respond to the secretagogue, histamine. Typical morphological changes from a resting to an acid-secreting active parietal cell were observed. In resting cultures of mouse parietal cells, the H + /K + ATPase displayed a cytoplasmic punctate staining pattern consistent with tubulovesicle element structures. Following histamine stimulation, an expansion of internal apical vacuole structures was observed together with a pronounced redistribution of the H + /K + ATPase from the cytoplasm to the apical vacuoles. A reproducible procedure to express genes of interest exogenously in these cultures of mouse parietal cells was also established. This method combines recombinant adenoviral transduction with magnetic field-assisted transfection resulting in ∼30% transduced parietal cells. Adenoviral-transduced parietal cells maintain their ability to undergo agonist-induced activation. This protocol will be useful for the isolation, culture and expression of genes in parietal cells from genetically modified mice and as such will be an invaluable tool for studying the complex exocytic and endocytic trafficking events of the H + /K + ATPase which underpin the regulation of acid secretion
-
Risk Mitigation in Preventing Adventitious Agent Contamination of Mammalian Cell Cultures.
Shiratori, Masaru; Kiss, Robert
2017-11-14
Industrial-scale mammalian cell culture processes have been contaminated by viruses during the culturing phase. Although the historical frequency of such events has been quite low, the impact of contamination can be significant for the manufacturing company and for the supply of the product to patients. This chapter discusses sources of adventitious agent contamination risk in a cell culture process, provides a semiquantitative assessment of such risks, and describes potential process barriers that can be used to reduce contamination risk. High-temperature, short-time (HTST) heat treatment is recommended as the process barrier of choice, when compatible with the process. A case study assessing the compatibility of HTST heat treatment with a cell culture medium is presented, and lessons learned are shared from our experiences over many years of developing and implementing virus barriers in mammalian cell culture processes. Graphical Abstract.
-
Yenuganti, Vengala Rao; Vanselow, Jens
2017-05-01
Cell culture models are essential for the detailed study of molecular processes. We analyze the dynamics of changes in a culture model of bovine granulosa cells. The cells were cultured for up to 8 days and analyzed for steroid production and gene expression. According to the expression of the marker genes CDH1, CDH2 and VIM, the cells maintained their mesenchymal character throughout the time of culture. In contrast, the levels of functionally important transcripts and of estradiol and progesterone production were rapidly down-regulated but showed a substantial up-regulation from day 4. FOXL2, a marker for granulosa cell identity, was also rapidly down-regulated after plating but completely recovered towards the end of culture. In contrast, expression of the Sertoli cell marker SOX9 and the lesion/inflammation marker PTGS2 increased during the first 2 days after plating but gradually decreased later on. We conclude that only long-term culture conditions (>4 days) allow the cells to recover from plating stress and to re-acquire characteristic granulosa cell features.
-
International Nuclear Information System (INIS)
Rooney, P.; Wright, E.G.
1993-01-01
Empty plastic tissue culture flasks were exposed to X-irradiation doses of 0.3-10.0 Gy, prior to the establishment of long-term bone marrow cultures. During the course of a 10 week culture period, all irradiated plastic flasks exhibited a dramatic decrease in the number of both haemopoietic stem cells and myeloid progenitor cells, in the non-adherent layer, when compared with controls. This decrease was not due to a decrease in the number of non-adherent cells produced. Histological examination of non-adherent cells showed an increase in mature granulocytic cells with few blast cells. Morphologically, the adherent layers of irradiated flasks demonstrated a delay in appearance or absence of fat cell production. X-irradiation of glass tissue culture flasks had no deleterious effect. (author)
-
Chhabra, Aastha; Jaiswal, Astha; Malhotra, Umang; Kohli, Shrey; Rani, Vibha
2012-09-01
In situ zymography is a unique technique for detection and localization of enzyme-substrate interactions majorly in histological sections. Substrate with quenched fluorogenic molecule is incorporated in gel over which tissue sections are mounted and then incubated in buffer. The enzymatic activity is observed in the form of fluorescent signal. With the advancements in the field of biological research, use of in vitro cell culture has become very popular and holds great significance in multiple fields including inflammation, cancer, stem cell biology and the still emerging 3-D cell cultures. The information on analysis of enzymatic activity in cell lines is inadequate presently. We propose a single-step methodology that is simple, sensitive, cost-effective, and functional to perform and study the 'in position' activity of enzyme on substrate for in vitro cell cultures. Quantification of enzymatic activity to carry out comparative studies on cells has also been illustrated. This technique can be applied to a variety of enzyme classes including proteases, amylases, xylanases, and cellulases in cell cultures.
-
Carney, Colleen M.; Muszynski, Jessica L.; Strotman, Lindsay N.; Lewis, Samantha R.; O'Connell, Rachel L.; Beebe, David J.; Theberge, Ashleigh B.; Jorgensen, Joan S.
2014-01-01
ABSTRACT Despite the fact that fetal Leydig cells are recognized as the primary source of androgens in male embryos, the mechanisms by which steroidogenesis occurs within the developing testis remain unclear. A genetic approach was used to visualize and isolate fetal Leydig cells from remaining cells within developing mouse testes. Cyp11a1-Cre mice were bred to mT/mG dual reporter mice to target membrane-tagged enhanced green fluorescent protein (GFP) within steroidogenic cells, whereas other cells expressed membrane-tagged tandem-dimer tomato red. Fetal Leydig cell identity was validated using double-labeled immunohistochemistry against GFP and the steroidogenic enzyme 3beta-HSD, and cells were successfully isolated as indicated by qPCR results from sorted cell populations. Because fetal Leydig cells must collaborate with neighboring cells to synthesize testosterone, we hypothesized that the fetal Leydig cell microenvironment defined their capacity for androgen production. Microfluidic culture devices were used to measure androstenedione and testosterone production of fetal Leydig cells that were cultured in cell-cell contact within a mixed population, were isolated but remained in medium contact via compartmentalized co-culture with other testicular cells, or were isolated and cultured alone. Results showed that fetal Leydig cells maintained their identity and steroidogenic activity for 3–5 days in primary culture. Microenvironment dictated proficiency of testosterone production. As expected, fetal Leydig cells produced androstenedione but not testosterone when cultured in isolation. More testosterone accumulated in medium from mixed cultures than from compartmentalized co-cultures initially; however, co-cultures maintained testosterone synthesis for a longer time. These data suggest that a combination of cell-cell contact and soluble factors constitute the ideal microenvironment for fetal Leydig cell activity in primary culture. PMID:25143354
-
Tailor-made three-dimensional hybrid scaffolds for cell cultures
International Nuclear Information System (INIS)
Psycharakis, Stylianos; Melissinaki, Vasileia; Giakoumaki, Anastasia; Ranella, Anthi; Tosca, Androniki
2011-01-01
The construction of the ideal three-dimensional scaffold for cell culture is one of the most intriguing topics in tissue engineering. It has been shown that cells can be cultured on most organic biomimetic materials, which now are losing popularity in favour of novel, hybrid systems. In this study, a series of photosensitive sol-gel hybrid materials, based on silicon-zirconium and silicon-titanium oxides, have been investigated for their suitability in three-dimensional scaffold fabrication. These materials can be structured by two-photon polymerization, a laser-based technique allowing the fabrication of micrometre-size structures with submicron resolution. The work presented here examined the effect of the organic/inorganic composition of the materials on cell behaviour and the establishment of a 'cell-culture friendly' environment. This is vital for cell adhesion, growth and differentiation, as the organic part of the material provides the soft matrix for cell growth, whereas the inorganic component gives the mechanical stability and rigidity of the three-dimensional structures. In addition, the use of femtosecond laser structuring permits the fabrication of a wide range of mechanically stable scaffolds of different sizes and shapes to be tested in terms of cell viability, proliferation and orientation.
-
Exploring culture, language and the perception of the nature of science
Sutherland, Dawn
2002-01-01
One dimension of early Canadian education is the attempt of the government to use the education system as an assimilative tool to integrate the First Nations and Me´tis people into Euro-Canadian society. Despite these attempts, many First Nations and Me´tis people retained their culture and their indigenous language. Few science educators have examined First Nations and Western scientific worldviews and the impact they may have on science learning. This study explored the views some First Nations (Cree) and Euro-Canadian Grade-7-level students in Manitoba had about the nature of science. Both qualitative (open-ended questions and interviews) and quantitative (a Likert-scale questionnaire) instruments were used to explore student views. A central hypothesis to this research programme is the possibility that the different world-views of two student populations, Cree and Euro-Canadian, are likely to influence their perceptions of science. This preliminary study explored a range of methodologies to probe the perceptions of the nature of science in these two student populations. It was found that the two cultural groups differed significantly between some of the tenets in a Nature of Scientific Knowledge Scale (NSKS). Cree students significantly differed from Euro-Canadian students on the developmental, testable and unified tenets of the nature of scientific knowledge scale. No significant differences were found in NSKS scores between language groups (Cree students who speak English in the home and those who speak English and Cree or Cree only). The differences found between language groups were primarily in the open-ended questions where preformulated responses were absent. Interviews about critical incidents provided more detailed accounts of the Cree students' perception of the nature of science. The implications of the findings of this study are discussed in relation to the challenges related to research methodology, further areas for investigation, science
-
Somatic embryogenesis in cell cultures of Glycine species.
Gamborg, O L; Davis, B P; Stahlhut, R W
1983-08-01
This report describes the development of procedures for the production of somatic embryos in cell cultures of Glycine species including soybean. The conditions for callus induction and initiation of rapidly growing cell suspension cultures were defined. Methods for inducing embryogenesis were tested on 16 lines of several Glycine species and cultivars of soybean. The SB-26 Culture of a G. soja gave the best results and was used in the experiments. Embryogenesis required the presence of picloram or 2,4-D. AMO 1618, CCC, PP-333 and Ancymidol enhanced the embryogenesis frequency. Plants of the G. soja (SB-26) were grown to maturity from seed-derived shoot tips. Characteristics of the plants are discussed.
-
Topological defects control collective dynamics in neural progenitor cell cultures
Kawaguchi, Kyogo; Kageyama, Ryoichiro; Sano, Masaki
2017-04-01
Cultured stem cells have become a standard platform not only for regenerative medicine and developmental biology but also for biophysical studies. Yet, the characterization of cultured stem cells at the level of morphology and of the macroscopic patterns resulting from cell-to-cell interactions remains largely qualitative. Here we report on the collective dynamics of cultured murine neural progenitor cells (NPCs), which are multipotent stem cells that give rise to cells in the central nervous system. At low densities, NPCs moved randomly in an amoeba-like fashion. However, NPCs at high density elongated and aligned their shapes with one another, gliding at relatively high velocities. Although the direction of motion of individual cells reversed stochastically along the axes of alignment, the cells were capable of forming an aligned pattern up to length scales similar to that of the migratory stream observed in the adult brain. The two-dimensional order of alignment within the culture showed a liquid-crystalline pattern containing interspersed topological defects with winding numbers of +1/2 and -1/2 (half-integer due to the nematic feature that arises from the head-tail symmetry of cell-to-cell interaction). We identified rapid cell accumulation at +1/2 defects and the formation of three-dimensional mounds. Imaging at the single-cell level around the defects allowed us to quantify the velocity field and the evolving cell density; cells not only concentrate at +1/2 defects, but also escape from -1/2 defects. We propose a generic mechanism for the instability in cell density around the defects that arises from the interplay between the anisotropic friction and the active force field.
-
Is cell culture a risky business? Risk analysis based on scientist survey data.
Shannon, Mark; Capes-Davis, Amanda; Eggington, Elaine; Georghiou, Ronnie; Huschtscha, Lily I; Moy, Elsa; Power, Melinda; Reddel, Roger R; Arthur, Jonathan W
2016-02-01
Cell culture is a technique that requires vigilance from the researcher. Common cell culture problems, including contamination with microorganisms or cells from other cultures, can place the reliability and reproducibility of cell culture work at risk. Here we use survey data, contributed by research scientists based in Australia and New Zealand, to assess common cell culture risks and how these risks are managed in practice. Respondents show that sharing of cell lines between laboratories continues to be widespread. Arrangements for mycoplasma and authentication testing are increasingly in place, although scientists are often uncertain how to perform authentication testing. Additional risks are identified for preparation of frozen stocks, storage and shipping. © 2015 UICC.
-
Cultural studies of science education
Higgins, Joanna; McDonald, Geraldine
2008-07-01
In response to Stetsenko's [2008, Cultural Studies of Science Education, 3] call for a more unified approach in sociocultural perspectives, this paper traces the origins of the use of sociocultural ideas in New Zealand from the 1970s to the present. Of those New Zealanders working from a sociocultural perspective who responded to our query most had encountered these ideas while overseas. More recently activity theory has been of interest and used in reports of work in early childhood, workplace change in the apple industry, and in-service teacher education. In all these projects the use of activity theory has been useful for understanding how the elements of a system can transform the activity. We end by agreeing with Stetsenko that there needs to be a more concerted approach by those working from a sociocultural perspective to recognise the contribution of others in the field.
-
The terminator syndrome: Science fiction, cinema and contemporary culture
Directory of Open Access Journals (Sweden)
J. Sey
1992-05-01
Full Text Available This paper examines the impact of contemporary technology on representations of the human body in American popular culture, focusing on James Cameron’s science fiction films The Terminator (1984 and The Terminator II - Judgment Day (1991 in both of which the key figures are cybernetic organisms (cyborgs or a robot which can exactly imitate the human form . The paper argues that the ability of modern film technology’ to represent the human form in robotic guise undercuts the distinction between nature and culture which maintains the position of the human being in society. The ability of the robot or cyborg to be ‘polygendered’ in particular, undermines the position of a properly oedipalized human body in society, one which balances the instinctual life against the rule of cultural law. As a result the second Terminator film attempts a recuperation of the category of the human by an oedipalization of the terminator cyborg.
-
Pigment Cell Differentiation in Sea Urchin Blastula-Derived Primary Cell Cultures
Ageenko, Natalya V.; Kiselev, Konstantin V.; Dmitrenok, Pavel S.; Odintsova, Nelly A.
2014-01-01
The quinone pigments of sea urchins, specifically echinochrome and spinochromes, are known for their effective antioxidant, antibacterial, antifungal, and antitumor activities. We developed in vitro technology for inducing pigment differentiation in cell culture. The intensification of the pigment differentiation was accompanied by a simultaneous decrease in cell proliferation. The number of pigment cells was two-fold higher in the cells cultivated in the coelomic fluids of injured sea urchins than in those intact. The possible roles of the specific components of the coelomic fluids in the pigment differentiation process and the quantitative measurement of the production of naphthoquinone pigments during cultivation were examined by MALDI and electrospray ionization mass spectrometry. Echinochrome A and spinochrome E were produced by the cultivated cells of the sand dollar Scaphechinus mirabilis in all tested media, while only spinochromes were found in the cultivated cells of another sea urchin, Strongylocentrotus intermedius. The expression of genes associated with the induction of pigment differentiation was increased in cells cultivated in the presence of shikimic acid, a precursor of naphthoquinone pigments. Our results should contribute to the development of new techniques in marine biotechnology, including the generation of cell cultures producing complex bioactive compounds with therapeutic potential. PMID:24979272
-
The Effect of Spaceflight on Bone Cell Cultures
Landis, William J.
1999-01-01
Understanding the response of bone to mechanical loading (unloading) is extremely important in defining the means of adaptation of the body to a variety of environmental conditions such as during heightened physical activity or in extended explorations of space or the sea floor. The mechanisms of the adaptive response of bone are not well defined, but undoubtedly they involve changes occurring at the cellular level of bone structure. This proposal has intended to examine the hypothesis that the loading (unloading) response of bone is mediated by specific cells through modifications of their activity cytoskeletal elements, and/or elaboration of their extracellular matrices. For this purpose, this laboratory has utilized the results of a number of previous studies defining molecular biological, biochemical, morphological, and ultrastructural events of the reproducible mineralization of a primary bone cell (osteoblast) culture system under normal loading (1G gravity level). These data and the culture system then were examined following the use of the cultures in two NASA shuttle flights, STS-59 and STS-63. The cells collected from each of the flights were compared to respective synchronous ground (1G) control cells examined as the flight samples were simultaneously analyzed and to other control cells maintained at 1G until the time of shuttle launch, at which point they were terminated and studied (defined as basal cells). Each of the cell cultures was assayed in terms of metabolic markers- gene expression; synthesis and secretion of collagen and non-collagenous proteins, including certain cytoskeletal components; assembly of collagen into macrostructural arrays- formation of mineral; and interaction of collagen and mineral crystals during calcification of the cultures. The work has utilized a combination of biochemical techniques (radiolabeling, electrophoresis, fluorography, Western and Northern Blotting, and light microscopic immunofluorescence) and structural
-
Culture of Mouse Neural Stem Cell Precursors
Currle, D. Spencer; Hu, Jia Sheng; Kolski-Andreaco, Aaron; Monuki, Edwin S.
2007-01-01
Primary neural stem cell cultures are useful for studying the mechanisms underlying central nervous system development. Stem cell research will increase our understanding of the nervous system and may allow us to develop treatments for currently incurable brain diseases and injuries. In addition, stem cells should be used for stem cell research aimed at the detailed study of mechanisms of neural differentiation and transdifferentiation and the genetic and environmental signals that direct the...
-
Induction of expression of two phenotypic markers of pulmonary type II cells in a cultured cell line
International Nuclear Information System (INIS)
Henderson, R.F.; Waide, J.J.; Scott, G.G.
1994-01-01
The functions of pulmonary type II cells, such as synthesis of pulmonary surfactant and metabolism of inhaled xenobiotics, can be studied in primary isolates of lung cells. However, isolated type II cells, when cultured, quickly lose the phenotypic expressions characteristics of type II cells, including surfactant lipid and protein synthesis and alkaline phosphatase (AP) activity. A cultured cell line that maintained expression of type II cell markers of differentiation would be advantageous for the study of such functions as surfactant synthesis and secretion. Such a cell line would allow generation of a large number of homogeneous cells for study. The purpose of the current study was to induce markers of differentiated type II cells in a cultured cell line to facilitate studies of factors that control surfactant synthesis and secretion
-
Li, Yi; Guo, Gang; Li, Li; Chen, Fei; Bao, Ji; Shi, Yu-Jun; Bu, Hong
2015-05-01
Mesenchymal stem cell (MSC) transplantation is a promising treatment of many diseases. However, conventional techniques with cells being cultured as a monolayer result in slow cell proliferation and insufficient yield to meet clinical demands. Three-dimensional (3D) culture systems are gaining attention with regard to recreating a complex microenvironment and to understanding the conditions experienced by cells. Our aim is to establish a novel 3D system for the culture of human umbilical cord MSCs (hUC-MSCs) within a real 3D microenvironment but with no digestion or passaging. Primary hUC-MSCs were isolated and grown in serum-free medium (SFM) on a suspension Rocker system. Cell characteristics including proliferation, phenotype and multipotency were recorded. The therapeutic effects of 3D-cultured hUC-MSCs on carbon tetrachloride (CCl4)-induced acute liver failure in mouse models were examined. In the 3D Rocker system, hUC-MSCs formed spheroids in SFM and maintained high viability and active proliferation. Compared with monolayer culture, the 3D-culture system yielded more hUC-MSCs cells within the same volume. The spheroids expressed higher levels of stem cell markers and displayed stronger multipotency. After transplantation into mouse, 3D hUC-MSCs significantly promoted the secretion of interferon-γ and interleukin-6 but inhibited that of tumor necrosis factor-α, thereby alleviating liver necrosis and promoting regeneration following CCl4 injury. The 3D culture of hUC-MSCs thus promotes cell yield and stemness maintenance and represents a promising strategy for hUC-MSCs expansion on an industrial scale with great potential for cell therapy and biotechnology.
-
An Exploration of Hispanic Mothers' Culturally Sustaining Experiences at an Informal Science Center
Weiland, Ingrid
2015-01-01
Science education reform focuses on learner-centered instruction within contexts that support learners' sociocultural experiences. The purpose of this study was to explore Hispanic mothers' experiences as accompanying adults at an informal science center within the context of culturally sustaining experiences, which include the fluidity…
-
DEFF Research Database (Denmark)
Claesson, Mogens Helweg; Ropke, C
1990-01-01
We show that thymic epithelial cells grown under serum-free conditions in a chemically defined culture medium can act as veto cells in vitro. The veto activity of thymic epithelial cells results in inactivation of specific alloreactive cytotoxic T-cell precursors at the clonal level. It is conclu......We show that thymic epithelial cells grown under serum-free conditions in a chemically defined culture medium can act as veto cells in vitro. The veto activity of thymic epithelial cells results in inactivation of specific alloreactive cytotoxic T-cell precursors at the clonal level...
-
Directory of Open Access Journals (Sweden)
Hensel Nancy
2005-04-01
Full Text Available Abstract In vitro stimulation of CD34+ cells with IL-2 induces NK cell differentiation. In order to define the stages of NK cell development, which influence their generation from CD34 cells, we cultured G-CSF mobilized peripheral blood CD34+ cells in the presence of stem cell factor and IL-2. After three weeks culture we found a diversity of CD56+ subsets which possessed granzyme A, but lacked the cytotoxic apparatus required for classical NK-like cytotoxicity. However, these CD56+ cells had the unusual property of inhibiting proliferation of K562 and P815 cell lines in a cell-contact dependent fashion.
-
Murry, Adam T.
Science has been identified as a crucial element in the competitiveness and sustainability of America in the global economy. American citizens, especially minority populations, however, are not pursuing science education or careers. Past research has implicated ‘attitudes toward science’ as an important factor in the public’s participation in science. I applied Ajzen’s (1991) Theory of Planned Behavior to attitudes toward science to predict science-related sustainability-action intentions and evaluated whether scientists and Native Americans differed in their general attitudes toward science, cultural values, and specific beliefs about science. Analyses revealed that positive attitude toward science and the cultural value of individualism predicted intentions to engage with science-related sustainability actions. Unexpectedly, scientists and Native Americans did not differ in their cultural values or positive attitude toward science. However, Natives Americans held significantly more negative attitude toward science than scientists. Implications for science education and attitudes towards science theory and application are discussed.
-
Cultured meat from stem cells: challenges and prospects.
Post, Mark J
2012-11-01
As one of the alternatives for livestock meat production, in vitro culturing of meat is currently studied. The generation of bio-artificial muscles from satellite cells has been ongoing for about 15 years, but has never been used for generation of meat, while it already is a great source of animal protein. In order to serve as a credible alternative to livestock meat, lab or factory grown meat should be efficiently produced and should mimic meat in all of its physical sensations, such as visual appearance, smell, texture and of course, taste. This is a formidable challenge even though all the technologies to create skeletal muscle and fat tissue have been developed and tested. The efficient culture of meat will primarily depend on culture conditions such as the source of medium and its composition. Protein synthesis by cultured skeletal muscle cells should further be maximized by finding the optimal combination of biochemical and physical conditions for the cells. Many of these variables are known, but their interactions are numerous and need to be mapped. This involves a systematic, if not systems, approach. Given the urgency of the problems that the meat industry is facing, this endeavor is worth undertaking. As an additional benefit, culturing meat may provide opportunities for production of novel and healthier products. Copyright © 2012 Elsevier Ltd. All rights reserved.
-
A carbon dioxide stripping model for mammalian cell culture in manufacturing scale bioreactors.
Xing, Zizhuo; Lewis, Amanda M; Borys, Michael C; Li, Zheng Jian
2017-06-01
Control of carbon dioxide within the optimum range is important in mammalian bioprocesses at the manufacturing scale in order to ensure robust cell growth, high protein yields, and consistent quality attributes. The majority of bioprocess development work is done in laboratory bioreactors, in which carbon dioxide levels are more easily controlled. Some challenges in carbon dioxide control can present themselves when cell culture processes are scaled up, because carbon dioxide accumulation is a common feature due to longer gas-residence time of mammalian cell culture in large scale bioreactors. A carbon dioxide stripping model can be used to better understand and optimize parameters that are critical to cell culture processes at the manufacturing scale. The prevailing carbon dioxide stripping models in literature depend on mass transfer coefficients and were applicable to cell culture processes with low cell density or at stationary/cell death phase. However, it was reported that gas bubbles are saturated with carbon dioxide before leaving the culture, which makes carbon dioxide stripping no longer depend on a mass transfer coefficient in the new generation cell culture processes characterized by longer exponential growth phase, higher peak viable cell densities, and higher specific production rate. Here, we present a new carbon dioxide stripping model for manufacturing scale bioreactors, which is independent of carbon dioxide mass transfer coefficient, but takes into account the gas-residence time and gas CO 2 saturation time. The model was verified by CHO cell culture processes with different peak viable cell densities (7 to 12 × 10 6 cells mL -1 ) for two products in 5,000-L and 25,000-L bioreactors. The model was also applied to a next generation cell culture process to optimize cell culture conditions and reduce carbon dioxide levels at manufacturing scale. The model provides a useful tool to understand and better control cell culture carbon dioxide
-
Culture of human cells in experimental units for spaceflight impacts on their behavior.
Cazzaniga, Alessandra; Moscheni, Claudia; Maier, Jeanette Am; Castiglioni, Sara
2017-05-01
Because space missions produce pathophysiological alterations such as cardiovascular disorders and bone demineralization which are very common on Earth, biomedical research in space is a frontier that holds important promises not only to counterbalance space-associated disorders in astronauts but also to ameliorate the health of Earth-bound population. Experiments in space are complex to design. Cells must be cultured in closed cell culture systems (from now defined experimental units (EUs)), which are biocompatible, functional, safe to minimize any potential hazard to the crew, and with a high degree of automation. Therefore, to perform experiments in orbit, it is relevant to know how closely culture in the EUs reflects cellular behavior under normal growth conditions. We compared the performances in these units of three different human cell types, which were recently space flown, i.e. bone mesenchymal stem cells, micro- and macrovascular endothelial cells. Endothelial cells are only slightly and transiently affected by culture in the EUs, whereas these devices accelerate mesenchymal stem cell reprogramming toward osteogenic differentiation, in part by increasing the amounts of reactive oxygen species. We conclude that cell culture conditions in the EUs do not exactly mimic what happens in a culture dish and that more efforts are necessary to optimize these devices for biomedical experiments in space. Impact statement Cell cultures represent valuable preclinical models to decipher pathogenic circuitries. This is true also for biomedical research in space. A lot has been learnt about cell adaptation and reaction from the experiments performed on many different cell types flown to space. Obviously, cell culture in space has to meet specific requirements for the safety of the crew and to comply with the unique environmental challenges. For these reasons, specific devices for cell culture in space have been developed. It is important to clarify whether these
-
Assessment of genetic and epigenetic variation during long-term Taxus cell culture.
Fu, Chunhua; Li, Liqin; Wu, Wenjuan; Li, Maoteng; Yu, Xiaoqing; Yu, Longjiang
2012-07-01
Gradual loss of secondary metabolite production is a common obstacle in the development of a large-scale plant cell production system. In this study, cell morphology, paclitaxel (Taxol®) biosynthetic ability, and genetic and epigenetic variations in the long-term culture of Taxus media cv Hicksii cells were assessed over a 5-year period to evaluate the mechanisms of the loss of secondary metabolites biosynthesis capacity in Taxus cell. The results revealed that morphological variations, gradual loss of paclitaxel yield and decreased transcriptional level of paclitaxel biosynthesis key genes occurred during long-term subculture. Genetic and epigenetic variations in these cultures were also studied at different times during culture using amplified fragment-length polymorphism (AFLP), methylation-sensitive amplified polymorphism (MSAP), and high-performance liquid chromatography (HPLC) analyses. A total of 32 primer combinations were used in AFLP amplification, and none of the AFLP loci were found to be polymorphic, thus no major genetic rearrangements were detected in any of the tested samples. However, results from both MSAP and HPLC indicated that there was a higher level of DNA methylation in the low-paclitaxel yielding cell line after long-term culture. Based on these results, we proposed that accumulation of paclitaxel in Taxus cell cultures might be regulated by DNA methylation. To our knowledge, this is the first report of increased methylation with the prolongation of culture time in Taxus cell culture. It provides substantial clues for exploring the gradual loss of the taxol biosynthesis capacity of Taxus cell lines during long-term subculture. DNA methylation maybe involved in the regulation of paclitaxel biosynthesis in Taxus cell culture.
-
Primary culture of human Schwann and schwannoma cells: improved and simplified protocol.
Dilwali, Sonam; Patel, Pratik B; Roberts, Daniel S; Basinsky, Gina M; Harris, Gordon J; Emerick, Kevin S; Stankovic, Konstantina M
2014-09-01
Primary culture of human Schwann cells (SCs) and vestibular schwannoma (VS) cells are invaluable tools to investigate SC physiology and VS pathobiology, and to devise effective pharmacotherapies against VS, which are sorely needed. However, existing culture protocols, in aiming to create robust, pure cultures, employ methods that can lead to loss of biological characteristics of the original cells, potentially resulting in misleading biological findings. We have developed a minimally manipulative method to culture primary human SC and VS cells, without the use of selective mitogens, toxins, or time-consuming and potentially transformative laboratory techniques. Schwann cell purity was quantified longitudinally using S100 staining in SC cultures derived from the great auricular nerve and VS cultures followed for 7 and 12 weeks, respectively. SC cultures retained approximately ≥85% purity for 2 weeks. VS cultures retained approximately ≥80% purity for the majority of the span of 12 weeks, with maximal purity of 87% at 2 weeks. The VS cultures showed high level of biological similarity (68% on average) to their respective parent tumors, as assessed using a protein array featuring 41 growth factors and receptors. Apoptosis rate in vitro negatively correlated with tumor volume. Our results, obtained using a faster, simplified culturing method than previously utilized, indicate that highly pure, primary human SC and VS cultures can be established with minimal manipulation, reaching maximal purity at 2 weeks of culture. The VS cultures recapitulate the parent tumors' biology to a great degree, making them relevant models to investigate VS pathobiology. Copyright © 2014 Elsevier B.V. All rights reserved.
-
Effects of viscoelastic ophthalmic solutions on cell cultures
Directory of Open Access Journals (Sweden)
Madhavan Hajib
1998-01-01
Full Text Available The development of mild but significant inflammation probably attributable to viscoelastic ophthalmic solutions in cataract surgery was recently brought to the notice of the authors, and hence a study of the effects of these solutions available in India, on cell cultures was undertaken. We studied the effects of 6 viscoelastic ophthalmic solutions (2 sodium hyaluronate designated as A and B, and 4 hydroxypropylmethylcellulose designated as C, D, E and F on HeLa, Vero and BHK-21 cell lines in tissue culture microtitre plates using undiluted, 1:10 and 1:100 dilutions of the solutions, and in cover slip cultures using undiluted solutions. Phase contrast microscopic examination of the solutions was also done to determine the presence of floating particles. The products D and F produced cytotoxic changes in HeLa cell line and these products also showed the presence of floating particles under phase contrast microscopy. Other products did not have any adverse effects on the cell lines nor did they show floating particles. The viscoelastic ophthalmic pharmaceutical products designated D and F have cytotoxic effects on HeLa cell line which appears to be a useful cell line for testing these products for their toxicity. The presence of particulate materials in products D and F indicates that the methods used for purification of the solution are not effective.
-
Free-energy carriers in human cultured muscle cells
Bolhuis, P. A.; de Zwart, H. J.; Ponne, N. J.; de Jong, J. M.
1985-01-01
Creatine phosphate (CrP), adenosine triphosphate (ATP), creatine kinase (CK), adenylate kinase (AK), protein, and DNA were quantified in human muscle cell cultures undergoing transition from dividing myoblasts to multinucleate myotubes. CrP is negligible in cultures grown in commonly applied media
-
Proteins associated with adaptation of cultured tobacco cells to NaCl
International Nuclear Information System (INIS)
Singh, N.K.; Handa, A.K.; Hasegawa, P.M.; Bressan, R.A.
1985-01-01
Cultured tobacco cells (Nicotiana tabacum L. cv Wisconsin 38) adapted to grow in medium containing high levels of NaCl or polyethylene glycol (PEG) produce several new or enhanced polypeptide bands on sodium dodecyl sulfate-polyarylamide gel electrophoresis. The intensities of some of the polypeptide bands increase with increasing levels of NaCl adaptation, while the intensities of other polypeptide bands are reduced. Synthesis of 26-kilodalton polypeptide(s) occurs at two different periods during culture growth of NaCl adapted cells. Unadapted cells also incorporate 35 S into a 26-kilodalton polypeptide during the later stage of culture growth beginning at midlog phase. The 26-kilodalton polypeptides from adapted and unadapted cells have similar partial proteolysis peptide maps and are immunologically cross-reactive. During adaptation to NaCl, unadapted cells synthesize and accumulate a major 26-kilodalton polypeptide, and the beginning of synthesis corresponds to the period of osmotic adjustment and culture growth. From their results, the authors suggest an involvement of the 26-kilodalton polypeptide in the adaptation of cultured tobacco cells to NaCl and water stress. 38 references, 11 figures, 2 tables
-
Duchenne muscular dystrophy: normal ATP turnover in cultured cells
International Nuclear Information System (INIS)
Fox, I.H.; Bertorini, T.; Palmieri, G.M.A.; Shefner, R.
1986-01-01
This paper examines ATP metabolism in cultured muscle cells and fibroblasts from patients with Duchenne dystrophy. ATP and ADP levels were the same in cultured cells from normal subjects and patients and there was no difference in ATP synthesis or degradation. The ATP synthesis was measured by the incorporation of C 14-U-adenine into aTP and ADP. although there was a significant decrease in radioactively labelled ATP after incubation with deoxyglucose in Duchenne muscle cells, there was no difference in ATP concentration of ADP metabolism
-
Kolokouri, Eleni; Plakitsi, Katerina
2012-01-01
This study uses history of science in teaching natural sciences from the early grades. The theoretical framework used is Cultural Historical Activity Theory (CHAT), which is a theory with expanding applications in different fields of science. The didactical scenario, in which history of science is used in a CHAT context, refers to Newton's…
-
Human dental pulp cell culture and cell transplantation with an alginate scaffold.
Kumabe, Shunji; Nakatsuka, Michiko; Kim, Gi-Seup; Jue, Seong-Suk; Aikawa, Fumiko; Shin, Je-Won; Iwai, Yasutomo
2006-02-01
Many studies on tissue stem cells have been conducted in the field of regenerative medicine, and some studies have indicated that cultured dental pulp mesenchymal cells secrete dentin matrix. In the present study we used alginate as a scaffold to transplant subcultured human dental pulp cells subcutaneously into the backs of nude mice. We found that when beta-glycerophosphate was added to the culture medium, dentin sialophosphoprotein mRNA coding dentin sialoprotein (DSP) was expressed. An increase in alkaline phosphatase, which is an early marker for odontoblast differentiation, was also demonstrated. At 6 weeks after implantation the subcutaneous formation of radio-opaque calcified bodies was observed in situ. Immunohistochemical and fine structure studies identified expression of type I collagen, type III collagen, and DSP in the mineralizing transplants. Isolated odontoblast-like cells initiated dentin-like hard tissue formation and scattered autolyzing apoptotic cells were also observed in the transplants. The study showed that subcultured dental pulp cells actively differentiate into odontoblast-like cells and induce calcification in an alginate scaffold.
-
Formation of a science of physical culture in Ukraine.
Directory of Open Access Journals (Sweden)
Timoshenko Ju.O.
2011-07-01
Full Text Available The process of Ukrainian physical culture science institutional development is researched, its historical particularities and trends are shown. The author used only the archive data. They helped to define the structure and quality stuff of scientific institution, social and sports problems which influenced the research. It is established that the appearance of the Ukrainian Research Institute of Physical Education has identified a new trend of Soviet life.
-
Microfluidic bioreactors for culture of non-adherent cells
DEFF Research Database (Denmark)
Shah, Pranjul Jaykumar; Vedarethinam, Indumathi; Kwasny, Dorota
2011-01-01
Microfluidic bioreactors (μBR) are becoming increasingly popular for cell culture, sample preparation and analysis in case of routine genetic and clinical diagnostics. We present a novel μBR for non-adherent cells designed to mimic in vivo perfusion of cells based on diffusion of media through...
-
Cytokines effects on radio-induced apoptosis in cortical and hippocampal rat cells in culture
International Nuclear Information System (INIS)
Coffigny, H.; Briot, D.; Le Nin, I.
2000-01-01
In the central nervous system in development the radio-induced cell death occurs mainly by apoptosis. The effects of modulating factors like cytokines were studied on this kind of death. To handle more easily parameters implicated in nerve cell apoptosis, we studied the effects of radiation with a in vitro system. Cells were isolated from rat foetal cortex and hippocampus, two of the major structures implicated in human mental retardation observed after exposition in utero at Hiroshima and Nagasaki. Cortical or hippocampal cells were isolated from 17 day-old rat foetuses by enzymatic and mechanical treatments and irradiated with 0.50 or 1 Gy. The cells from both structures were cultured 1 or 3 days in serum free medium. Cytokines like βNGF, NT3, EGF, βTGF, α and βFGF, IGF I and II, interleukines like Il 1β, Il 2 and IL 6 were added to the medium. In 3 days cortical cell culture, only βFGF increased cell survival with as little as 10 ng/ml. This effect was dose dependent. In hippocampal cell culture, no significant increase of cell survival occurred with 10 ng/ml of any cytokines. In the same system culture with 1 Gy irradiation, the positive or negative effect of the association of βFGF with another cytokine was tested on cell survival. Only the association with EGF induced higher cell survival in cortical cell culture. In hippocampal cell culture where βFGF alone had no effect, the cell survival was not modified by the association. In the same system, the triple association of βFGF-EGF with another cytokine was tested on hippocampal and cortical cell cultures. No significant effect was observed in both cultures but cell survival trented to decrease with βTGF. In order to avoid the mitotic effect of cytokines in the 3 day-old culture, experiments were carried out on 20 hours cell culture, before the end of the first round of the cell cycle, with the selected cytokines (βFGF or βFGF-EGF). Without irradiation, the percentage of cortical cell survival
-
International Nuclear Information System (INIS)
Lee, Jingu; Park, Sangkyu; Roh, Sangho
2015-01-01
A loss of salivary gland function often occurs after radiation therapy in head and neck tumors, though secretion of saliva by the salivary glands is essential for the health and maintenance of the oral environment. Transplantation of salivary acinar cells (ACs), in part, may overcome the side effects of therapy. Here we directly differentiated mouse adipose-derived stromal cells (ADSCs) into ACs using a co-culture system. Multipotent ADSCs can be easily collected from stromal vascular fractions of adipose tissues. The isolated ADSCs showed positive expression of markers such as integrin beta-1 (CD29), cell surface glycoprotein (CD44), endoglin (CD105), and Nanog. The cells were able to differentiate into adipocytes, osteoblasts, and neural-like cells after 14 days in culture. ADSCs at passage 2 were co-cultured with mouse ACs in AC culture medium using the double-chamber (co-culture system) to avoid mixing the cell types. The ADSCs in this co-culture system expressed markers of ACs, such as α-amylases and aquaporin5, in both mRNA and protein. ADSCs cultured in AC-conditioned medium also expressed AC markers. Cellular proliferation and senescence analyses demonstrated that cells in the co-culture group showed lower senescence and a higher proliferation rate than the AC-conditioned medium group at Days 14 and 21. The results above imply direct conversion of ADSCs into ACs under the co-culture system; therefore, ADSCs may be a stem cell source for the therapy for salivary gland damage. - Highlights: • ADSCs could transdifferentiate into acinar cells (ACs) using ACs co-culture (CCA). • Transdifferentiated ADSCs expressed ACs markers such as α-amylase and aquaporin5. • High proliferation and low senescence were presented in CCA at Day 14. • Transdifferentiation of ADSCs into ACs using CCA may be an appropriate method for cell-based therapy
-
Energy Technology Data Exchange (ETDEWEB)
Lee, Jingu; Park, Sangkyu; Roh, Sangho, E-mail: sangho@snu.ac.kr
2015-05-15
A loss of salivary gland function often occurs after radiation therapy in head and neck tumors, though secretion of saliva by the salivary glands is essential for the health and maintenance of the oral environment. Transplantation of salivary acinar cells (ACs), in part, may overcome the side effects of therapy. Here we directly differentiated mouse adipose-derived stromal cells (ADSCs) into ACs using a co-culture system. Multipotent ADSCs can be easily collected from stromal vascular fractions of adipose tissues. The isolated ADSCs showed positive expression of markers such as integrin beta-1 (CD29), cell surface glycoprotein (CD44), endoglin (CD105), and Nanog. The cells were able to differentiate into adipocytes, osteoblasts, and neural-like cells after 14 days in culture. ADSCs at passage 2 were co-cultured with mouse ACs in AC culture medium using the double-chamber (co-culture system) to avoid mixing the cell types. The ADSCs in this co-culture system expressed markers of ACs, such as α-amylases and aquaporin5, in both mRNA and protein. ADSCs cultured in AC-conditioned medium also expressed AC markers. Cellular proliferation and senescence analyses demonstrated that cells in the co-culture group showed lower senescence and a higher proliferation rate than the AC-conditioned medium group at Days 14 and 21. The results above imply direct conversion of ADSCs into ACs under the co-culture system; therefore, ADSCs may be a stem cell source for the therapy for salivary gland damage. - Highlights: • ADSCs could transdifferentiate into acinar cells (ACs) using ACs co-culture (CCA). • Transdifferentiated ADSCs expressed ACs markers such as α-amylase and aquaporin5. • High proliferation and low senescence were presented in CCA at Day 14. • Transdifferentiation of ADSCs into ACs using CCA may be an appropriate method for cell-based therapy.
-
Universal lab-on-a-chip platform for complex, perfused 3D cell cultures
Sonntag, F.; Schmieder, F.; Ströbel, J.; Grünzner, S.; Busek, M.; Günther, K.; Steege, T.; Polk, C.; Klotzbach, U.
2016-03-01
The miniaturization, rapid prototyping and automation of lab-on-a-chip technology play nowadays a very important role. Lab-on-a-chip technology is successfully implemented not only for environmental analysis and medical diagnostics, but also as replacement of animals used for the testing of substances in the pharmaceutical and cosmetics industries. For that purpose the Fraunhofer IWS and partners developed a lab-on-a-chip platform for perfused cell-based assays in the last years, which includes different micropumps, valves, channels, reservoirs and customized cell culture modules. This technology is already implemented for the characterization of different human cell cultures and organoids, like skin, liver, endothelium, hair follicle and nephron. The advanced universal lab-on-a-chip platform for complex, perfused 3D cell cultures is divided into a multilayer basic chip with integrated micropump and application-specific 3D printed cell culture modules. Moreover a technology for surface modification of the printed cell culture modules by laser micro structuring and a complex and flexibly programmable controlling device based on an embedded Linux system was developed. A universal lab-on-a-chip platform with an optional oxygenator and a cell culture module for cubic scaffolds as well as first cell culture experiments within the cell culture device will be presented. The module is designed for direct interaction with robotic dispenser systems. This offers the opportunity to combine direct organ printing of cells and scaffolds with the microfluidic cell culture module. The characterization of the developed system was done by means of Micro-Particle Image Velocimetry (μPIV) and an optical oxygen measuring system.
-
Polymer microfilters with nanostructured surfaces for the culture of circulating cancer cells
International Nuclear Information System (INIS)
Makarova, Olga V.; Adams, Daniel L.; Divan, Ralu; Rosenmann, Daniel; Zhu, Peixuan; Li, Shuhong; Amstutz, Platte; Tang, Cha-Mei
2016-01-01
There is a critical need to improve the accuracy of drug screening and testing through the development of in vitro culture systems that more effectively mimic the in vivo environment. Surface topographical features on the nanoscale level, in short nanotopography, effect the cell growth patterns, and hence affect cell function in culture. We report the preliminary results on the fabrication, and subsequent cellular growth, of nanoscale surface topography on polymer microfilters using cell lines as a precursor to circulating tumor cells (CTCs). To create various nanoscale features on the microfilter surface, we used reactive ion etching (RIE) with and without an etching mask. An anodized aluminum oxide (AAO) membrane fabricated directly on the polymer surface served as an etching mask. Polymer filters with a variety of modified surfaces were used to compare the effects on the culture of cancer cell lines in blank culture wells, with untreated microfilters or with RIE-treated microfilters. We then report the differences of cell shape, phenotype and growth patterns of bladder and glioblastoma cancer cell lines after isolation on the various types of material modifications. Our data suggest that RIE modified polymer filters can isolate model cell lines while retaining ell viability, and that the RIE filter modification allows T24 monolayering cells to proliferate as a structured cluster. - Highlights: • Surface topographical effects the growth patterns and cell function of cancer cells • Nanoscale surface topography on polymer filters for circulating tumor cell culture • Membrane fabricated directly on polymer surfaces utilized for polymer etching • Nanotopography alters cell shape, phenotype and growth patterns of cancer cells • Nanoscale surface topography dictates monolayering or 3D structured cell culture
-
Polymer microfilters with nanostructured surfaces for the culture of circulating cancer cells
Energy Technology Data Exchange (ETDEWEB)
Makarova, Olga V. [Creatv MicroTech, Inc., 2242 West Harrison St., Chicago 60612, IL (United States); Adams, Daniel L., E-mail: dan@creatvmicrotech.com [Creatv MicroTech, Inc., 1 Deer Park Drive, Monmouth Junction, NJ 08852 (United States); Divan, Ralu; Rosenmann, Daniel [Center for Nanoscale Materials, Argonne National Laboratory, 9700 South Cass Ave., Argonne 60439, IL (United States); Zhu, Peixuan; Li, Shuhong; Amstutz, Platte; Tang, Cha-Mei [Creatv MicroTech, Inc., 11609 Lake Potomac Drive, Potomac 20854, MD (United States)
2016-09-01
There is a critical need to improve the accuracy of drug screening and testing through the development of in vitro culture systems that more effectively mimic the in vivo environment. Surface topographical features on the nanoscale level, in short nanotopography, effect the cell growth patterns, and hence affect cell function in culture. We report the preliminary results on the fabrication, and subsequent cellular growth, of nanoscale surface topography on polymer microfilters using cell lines as a precursor to circulating tumor cells (CTCs). To create various nanoscale features on the microfilter surface, we used reactive ion etching (RIE) with and without an etching mask. An anodized aluminum oxide (AAO) membrane fabricated directly on the polymer surface served as an etching mask. Polymer filters with a variety of modified surfaces were used to compare the effects on the culture of cancer cell lines in blank culture wells, with untreated microfilters or with RIE-treated microfilters. We then report the differences of cell shape, phenotype and growth patterns of bladder and glioblastoma cancer cell lines after isolation on the various types of material modifications. Our data suggest that RIE modified polymer filters can isolate model cell lines while retaining ell viability, and that the RIE filter modification allows T24 monolayering cells to proliferate as a structured cluster. - Highlights: • Surface topographical effects the growth patterns and cell function of cancer cells • Nanoscale surface topography on polymer filters for circulating tumor cell culture • Membrane fabricated directly on polymer surfaces utilized for polymer etching • Nanotopography alters cell shape, phenotype and growth patterns of cancer cells • Nanoscale surface topography dictates monolayering or 3D structured cell culture.
-
(De)colonizing culture in community psychology: reflections from critical social science.
Reyes Cruz, Mariolga; Sonn, Christopher C
2011-03-01
Since its inception, community psychology has been interested in cultural matters relating to issues of diversity and marginalization. However, the field has tended to understand culture as static social markers or as the background for understanding group differences. In this article the authors contend that culture is inseparable from who we are and what we do as social beings. Moreover, culture is continually shaped by socio-historical and political processes intertwined within the globalized history of power. The authors propose a decolonizing standpoint grounded in critical social science to disrupt understandings of cultural matters that marginalize others. This standpoint would move the field toward deeper critical thinking, reflexivity and emancipatory action. The authors present their work to illustrate how they integrate a decolonizing standpoint to community psychology research and teaching. They conclude that community psychology must aim towards intercultural work engaging its political nature from a place of ontological/epistemological/methodological parity.
-
Transformation of conceptual basis of political science under cultural and historical context
Directory of Open Access Journals (Sweden)
O. S. Tokovenko
2017-07-01
Full Text Available The paper is submitted to a scientific discussion the possibility of considering the idea of political science, based on the criteria of intellectual integrity and disciplinary unity. In this context, generally accepted among professionals and political scientists idea that political science as a scientific discipline occurs in the early twentieth century and its conceptual framework is still in a state of development, and a long preceding period should be characterized as a period of political thought is being challenged. The main idea that is being proved is recognition the existence of such scientific discipline as political science requires recognition of the existence of specific inherent ideals of science, cognitive standards, rules, procedures, explanations, etc. They allow political thinkers from the ancient world as well as modern researchers to combine it into a single, unique, different from others in their methodological principles and heuristic potential Science. It is convinced that the existence of intellectual integrity and disciplinary unity in Political Science is possible due to the existence of the ideals of scholarship, which are closely related to the cultural and historical context in which Political Science is being developed. The possibility of applying such disciplinary and integrated approach is considered as an example of the impact that was made by changes of the Great French Revolution and its consequences on transformation of the conceptual framework of Political Science. It is concluded that the consideration of the peculiarities of political thought development in the social and cultural contexts related to the events of the Great French Revolution and its consequences argues that political science is responsive to changing the social context, makes changes in categorical apparatus, introduces the new field of scientific inquiry, actualized subject field. These actions are due to the specific disciplinary unity
-
Towards a Cultural Psychology of Science:Economics and Economists in Contemporary Chile
Marco Carre Benzi, David
2017-01-01
The present thesis is an enquiry about two distinct but complementary issues: the personal dimension of scientific activity, and the influential role that economists have had during the last decades in Chile. Regarding the former, this work complements existing philosophical, social, and psychological studies of science with a cultural psychology perspective. This perspective aims to be sensitive to the personal nature of the scientific activity but also to the cultural conditions in which sc...
-
Directory of Open Access Journals (Sweden)
AA Pourfatollah
2005-10-01
Full Text Available Introduction: Considering the importance of co-culture in differentiation of embryonic stem cells, the aim of this study was evaluation of the effect of co-culturing fetal liver stroma cells with P19 cells on the line of differentiation. Materials and Methods: For this purpose, P19 cells were cultured directly in semisolid medium. These cells proliferated and primarily differentiated to colonies know as embryoid bodies (EBs after 8-12 days. The Ebs cells were trypsinized and dissociated to single or double cells. Then these cells were co-cultured on the mouse fetal liver feeder layer in the absence of exogenous factors. After 14-18 days, the colonies were studied morphologically by benzidine and giemsa staining and also counted under invert microscope. Results: The percentages of benzidine positive (or erythroid and negative colonies were 94% and 6% respectively and also the cells of colonies were studied by Giemsa staining. Results showed that they were myeloid or lymphoid type cells. Thus, the results show that in the presence of mouse fetal liver feeder layer, the number of erythroid colonies was increased. Conclusions: Therefore, this technique may be effective for differentiation of stem cells from different sources into hematopoietic cells and can be used in future for human cell therapy.
-
Directory of Open Access Journals (Sweden)
Arno Amann
Full Text Available INTRODUCTION: We describe a novel 3D co-culture model using non-small cell lung cancer (NSCLC cell lines in combination with lung fibroblasts. This model allows the investigation of tumour-stroma interactions and addresses the importance of having a more in vivo like cell culture model. METHODS: Automation-compatible multi-well hanging drop microtiter plates were used for the production of 3D mono- and co-cultures. In these hanging drops the two NSCLC cell lines A549 and Colo699 were cultivated either alone or co-cultured with lung fibroblasts. The viability of tumour spheroids was confirmed after five and ten days by using Annexin V/Propidium Iodide staining for flow-cytometry. Tumour fibroblast spheroid formation was characterized by scanning electron microscope (SEM, semi-thin sections, fluorescence microscope and immunohistochemistry (IHC. In addition to conventional histology, protein expression of E-Cadherin, vimentin, Ki67, fibronectin, cytokeratin 7 and α-smooth muscle actin (α-SMA was investigated by IHC. RESULTS: Lower viability was observed in A549 monocultures compared to co-cultures, whereas Colo699 monocultures showed better viability compared to co-cultures. Ki67 expression varied significantly between mono- and co-cultures in both tumour cell lines. An increase of vimentin and decreased E-Cadherin expression could be detected during the course of the cultivation suggesting a transition to a more mesenchymal phenotype. Furthermore, the fibroblast cell line showed an expression of α-SMA only in co-culture with the cancer cell line A549, thereby indicating a mesenchymal to mesenchymal shift to an even more myofibroblast phenotype. CONCLUSION: We demonstrate that our method is a promising tool for the generation of tumour spheroid co-cultures. Furthermore, these spheroids allow the investigation of tumour-stroma interactions and a better reflection of in vivo conditions of cancer cells in their microenvironment. Our method holds
-
Cell culture plastics with immobilized interleukin-4 for monocyte differentiation
DEFF Research Database (Denmark)
Hansen, Morten; Hjortø, Gertrud Malene; Met, Özcan
2011-01-01
Standard cell culture plastic was surface modified by passive adsorption or covalent attachment of interleukin (IL)-4 and investigated for its ability to induce differentiation of human monocytes into mature dendritic cells, a process dose-dependently regulated by IL-4. Covalent attachment of IL-4...... in water instead of phosphate-buffered saline. Passively adsorbed IL-4 was observed to induce differentiation to dendritic cells, but analysis of cell culture supernatants revealed that leakage of IL-4 into solution could account for the differentiation observed. Covalent attachment resulted in bound IL-4...... at similar concentrations to the passive adsorption process, as measured by enzyme-linked immunosorbent assays, and the bound IL-4 did not leak into solution to any measurable extent during cell culture. However, covalently bound IL-4 was incapable of inducing monocyte differentiation. This may be caused...
-
A microfluidic cell culture device with integrated microelectrodes for barrier studies
DEFF Research Database (Denmark)
Tan, Hsih-Yin; Dufva, Martin; Kutter, Jörg P.
We present an eight cell culture microfluidic device fabricated using thiol-ene ‘click’ chemistry with embedded microelectrodes for evaluating barrier properties of human intestinal epithelial cells. The capability of the microelectrodes for trans-epithelial electrical resistance (TEER) measureme......) measurements was demonstrated by using confluent human colorectal epithelial cells (Caco-2) and rat fibroblast (CT 26) cells cultured in the microfluidic device....
-
Ferguson, Joseph Paul; Kameniar, Barbara
2014-10-01
This paper investigates the cognitive experiences of four religious students studying evolutionary biology in an inner city government secondary school in Melbourne, Australia. The participants in the study were identified using the Religious Background and Behaviours questionnaire (Connors, Tonigan, & Miller, 1996). Participants were interviewed and asked to respond to questions about their cognitive experiences of studying evolutionary biology. Students' responses were analysed using cultural analysis of discourse to construct a cultural model of religious students of science. This cultural model suggests that these students employ a human schema and a non-human schema, which assert that humans are fundamentally different from non-humans in terms of origins and that humans have a transcendental purpose in life. For these students, these maxims seem to be challenged by their belief that evolutionary biology is dictated by metaphysical naturalism. The model suggests that because the existential foundation of these students is challenged, they employ a believing schema to classify their religious explanations and a learning schema to classify evolutionary biology. These schemas are then hierarchically arranged with the learning schema being made subordinate to the believing schema. Importantly, these students are thus able to maintain their existential foundation while fulfilling the requirements of school science. However, the quality of this "learning" is questionable.
-
Two cultures are better than one: Earth sciences and Art for a better planet sustainability
Lanza, Tiziana; Rubbia, Giuliana; Negrete, Aquiles
2015-04-01
Climate change, pollution, desertification, natural hazard, animals' extinction are some of the problems we face every day. Very often Science and Technology are charged of the solutions while Art is intended mainly for entertainment. Are we sure this is the right attitude? "Technology is a queer thing. It brings you gifts with one hand, and stabs you in the back with the other", says C.P.Snow, author of a milestone book on the Two Cultures, namely Sciences and Humanities. If Science can drive to a rigorous knowledge of the Earth speaking to people's mind, Technology is Science in action. When individuals act very often the reasons behind their actions are linked to their education, values, sense of beauty, presence or absence of feelings, all things pertaining to the emotional sphere of humans usually addressed by humanistic culture. But if in one hand, Science and Technology cannot be left alone to solve the impelling problems that are deteriorating not only our planet resources but also our quality of life, on the other hand the humanistic culture can find a powerful ally in scientific culture for re-awakening in everybody the sense of beauty, values and respect for the planet. To know Earth is to love Earth, since nature is in itself a work of Art. Earth sciences dig out all the secrets that make our planet a unique place in the Universe we know. Every single phenomena can be seen then in a double face value. An Aurora, for instance, can inspire poetry for its beauty and colors but always remains the result of the interaction between the solar wind and the Earth magnetic field. And, most important, an Aurora will never inspire negative feelings. To make our part in creating a common field between Art and Earth sciences, we have created a blog and a related FaceBook page to collect, browsing the web, all the experiences in this trend, to find out that many scientists and artists are already working in this direction as a final and enjoyable surprise.
-
A study of chromosomal aberrations in amniotic fluid cell cultures.
Wolstenholme, J; Crocker, M; Jonasson, J
1988-06-01
This paper represents the analysis of 1916 routine amniotic fluid specimens harvested by an in situ fixation technique in a prospective study with regard to cultural chromosome anomalies. Excluding constitutional abnormalities, 2.9 per cent of 19,432 cells analysed showed some form of chromosome anomaly, terminal deletions (57 per cent) and chromatid/chromosome breaks and gaps (18 per cent) being the most frequent, followed by interchange aberrations (13 per cent) and trisomy (5 per cent). No case was found of more than one colony from the same culture showing the same anomaly without it being present in other cultures from the same fluid. The wholly abnormal colonies had a surplus of trisomies and from the mathematical considerations presented one may infer that these are likely to reflect the presence of abnormal cells in the amniotic fluid. Partly abnormal colonies appeared at a frequency that would correspond to virtual absence of selection against chromosomally abnormal cells when cultured in vitro. The aberrations found were similar to those seen as single cell anomalies, except for chromatid breaks and exchanges. The data suggest a basic preferential induction of trisomy for chromosomes 2, 18, 21, and the Y-chromosome. Structural aberrations showed a marked clustering of breakpoints around the centromeres. The frequency of mutant cells was low (1.4 X 10(-3)) before culture was initiated. At harvest, the frequency of abnormal cells was much higher (3 X 10(-2)) corresponding to 3 X 10(-3) mutations per cell per generation accumulating over approximately ten generations in vitro.
-
Directory of Open Access Journals (Sweden)
Jian Ning
2011-01-01
Full Text Available Most anti-cancer immunotherapeutic strategies involving dendritic cells (DC as vaccines rely upon the adoptive transfer of DC loaded with exogenous tumour-peptides. This study utilized human acute myeloid leukemia (AML cells as progenitors from which functional dendritic-like antigen presenting cells (DLC were generated, that constitutively express tumour antigens for recognition by CD8+ T cells. DLC were generated from AML cell lines KG-1 and MUTZ-3 using rapid culture techniques and appropriate cytokines. DLC were evaluated for their cell-surface phenotype, antigen uptake and ability to stimulate allogeneic responder cell proliferation, and production of IFN-γ; compared with DC derived from normal human PBMC donors. KG-1 and MUTZ-3 DLC increased expression of CD80, CD83, CD86, and HLA-DR, and MUTZ-3 DLC downregulated CD14 and expressed CD1a. Importantly, both KG-1 and MUTZ-3-derived DLC promoted proliferation of allogeneic responder cells more efficiently than unmodified cells; neither cells incorporated FITC-labeled dextran, but both stimulated IFN-γ production from responding allogeneic CD8+ T cells. Control DC produced from PBMC using the FastDC culture also expressed high levels of critical cell surface ligands and demonstrated good APC function. This paper indicates that functional DLC can be cultured from the AML cell lines KG-1 and MUTZ-3, and FastDC culture generates functional KG-1 DLC.
-
[Culture of pancreatic progenitor cells in hanging drop and on floating filter].
Ma, Feng-xia; Chen, Fang; Chi, Ying; Yang, Shao-guang; Lu, Shi-hong; Han, Zhong-chao
2013-06-01
To construct a method to culture pancreatic progenitor cells in hanging drop and on floating filter,and to examine if pancreatic progenitor cells can differentiate into mature endocrine cells with this method. Murine embryos at day 12.5 were isolated and digested into single cells,which were then cultured in hanging drop for 24h and formed spheres.Spheres were cultured on the filter for 6 days,which floated in the dish containing medium.During culture,the expressions of pancreas duodenum homeobox-1(PDX-1)and neurogenin3(Ngn3)were determined.The expressions of endocrine and exocrine markers,insulin,glucagon,and carboxypeptidase(CPA)were determined on day 7 by immunohistochemistry.Insulin secretion of spheres stimulated by glucose was detected by ELISA.The changes of pancreatic marker expressions during culture were monitored by real-time polymerase chain reaction(PCR). One day after the culture,there were still a large amount of PDX-1 positive cells in pancreatic spheres,and these cells proliferated.On day 3,high expression of Ngn3 was detected,and the Ngn3-positive cells did not proliferate.On day 7,The expressions of endocrine and exocrine markers in the differentiated pancreatic progenitor cells were detected,which were consistent with that in vivo.Insulin was secreted by spheres upon the stimulation of glucose. In hanging drop and on floating filter,pancreatic progenitor cells can differentiate into mature endocrine cells.
-
A co-cultured skin model based on cell support membranes
International Nuclear Information System (INIS)
Dai, N.-T.; Yeh, M.-K.; Liu, Demeral David; Adams, E.F.; Chiang, C.-H.; Yen, C.-Y.; Shih, C.-M.; Sytwu, H.-K.; Chen, Tim-Mo; Wang, H.-J.; Williamson, M.R.; Coombes, A.G.A.
2005-01-01
Tissue engineering of skin based on collagen: PCL biocomposites using a designed co-culture system is reported. The collagen: PCL biocomposites having collagen: PCL (w/w) ratios of 1:4, 1:8, and 1:20 have been proven to be biocompatible materials to support both adult normal human epidermal Keratinocyte (NHEK) and mouse 3T3 fibroblast growth in cell culture, respectively, by Dai, Coombes, et al. in 2004. Films of collagen: PCL biocomposites were prepared using non-crosslinking method by impregnation of lyophilized collagen mats with PCL/dichloromethane solutions followed by solvent evaporation. To mimic the dermal/epidermal structure of skin, the 1:20 collagen: PCL biocomposites were selected for a feasibility study of a designed co-culture technique that would subsequently be used for preparing fibroblast/biocomposite/keratinocyte skin models. A 55.3% increase in cell number was measured in the designed co-culture system when fibroblasts were seeded on both sides of a biocomposite film compared with cell culture on one surface of the biocomposite in the feasibility study. The co-culture of human keratinocytes and 3T3 fibroblasts on each side of the membrane was therefore studied using the same co-culture system by growing keratinocytes on the top surface of membrane for 3 days and 3T3 fibroblasts underneath the membrane for 6 days. Scanning electron microscopy (SEM) and immunohistochemistry assay revealed good cell attachment and proliferation of both human keratinocytes and 3T3 fibroblasts with these two types of cells isolated well on each side of the membrane. Using a modified co-culture technique, a co-cultured skin model presenting a confluent epidermal sheet on one side of the biocomposite film and fibroblasts populated on the other side of the film was developed successfully in co-culture system for 28 days under investigations by SEM and immunohistochemistry assay. Thus, the design of a co-culture system based on 1:20 (w/w) collagen: PCL biocomposite
-
Micro fluidic System for Culturing and Monitoring of Neuronal Cells and Tissue
DEFF Research Database (Denmark)
Bakmand, Tanya; Waagepetersen, Helle S.
The aim of this Ph.D. project was to combine experience within cell and tissue culturing, electrochemistry and microfabrication in order to develop an in vivo-like fluidic culturing platform, challenging the traditional culturing methods. The first goal was to develope a fluidic system for cultur...... with mass production. The last part of this thesis also includes perspectives on how to expand the latest designed device to facilitate culturing of tissue and co-culturing of cells....
-
Proliferation differentiation and therapeutic effect of short-term cultured murine bone marrow cells
International Nuclear Information System (INIS)
Zhao Zekun; Cong Jianbo
1986-01-01
Murine bone marrow cells were cultured in conditioned medium of muscle. After 24 hours of culture, both adherent and suspended cells appeared in the culture. The adherent cells mainly consisted of macrophages and the suspended cells were predominantly granulocytes. After 6 days, the total number of nucleated cells and CFU-C in the culture increased about 400% and 600% respectively, but CFU-S reduced to 21% approximately. Lymphocytes persisted only for 4 days. The stem cells (CFU-S) from 6-day culture were injected into the lethally irradiated syngenic mice. The 30 day survival rate of the treated mice was 89% whereas that of the controls was only 7%. The bone marrow cells in 2/8 of recipients sacrificed at 30 or 60 days were of donor type and 6/8 of the recipients were chimeras
-
Siemensma, André; Babcock, James; Wilcox, Chris; Huttinga, Hans
In the light of the growing demand for high quality plant-derived hydrolysates (i.e., HyPep™ and UltraPep™ series), Sheffield Bio-Science has developed a new hydrolysate platform that addresses the need for animal-free cell culture medium supplements while also minimizing variability concerns. The platform is based upon a novel approach to enzymatic digestion and more refined processing. At the heart of the platform is a rationally designed animal component-free (ACF) enzyme cocktail that includes both proteases and non-proteolytic enzymes (hydrolases) whose activities can also liberate primary components of the polymerized non-protein portion of the raw material. This enzyme system is added during a highly optimized process step that targets specific enzyme-substrate reactions to expand the range of beneficial nutritional factors made available to cells in culture. Such factors are fundamental to improving the bio-performance of the culture system, as they provide not merely growth-promoting peptides and amino acids, but also key carbohydrates, lipids, minerals, and vitamins that improve both rate and quality of protein expression, and serve to improve culture life due to osmo-protectant and anti-apoptotic properties. Also of significant note is that, compared to typical hydrolysates, the production process is greatly reduced and requires fewer steps, intrinsically yielding a better-controlled and therefore more reproducible product. Finally, the more sophisticated approach to enzymatic digestion renders hydrolysates more amenable to sterile filtration, allowing hydrolysate end users to experience streamlined media preparation and bioreactor supplementation activities. Current and future development activities will evolve from a better understanding of the complex interactions within a handful of key biochemical pathways that impact the growth and productivity of industrially relevant organisms. Presented in this chapter are some examples of the efforts that
-
A Novel Microgravity Simulator Applicable for Three-Dimensional Cell Culturing
Wuest, Simon L.; Richard, Stéphane; Walther, Isabelle; Furrer, Reinhard; Anderegg, Roland; Sekler, Jörg; Egli, Marcel
2014-10-01
Random Positioning Machines (RPM) were introduced decades ago to simulate microgravity. Since then numerous experiments have been carried out to study its influence on biological samples. The machine is valued by the scientific community involved in space relevant topics as an excellent experimental tool to conduct pre-studies, for example, before sending samples into space. We have developed a novel version of the traditional RPM to broaden its operative range. This novel version has now become interesting to researchers who are working in the field of tissue engineering, particularly those interested in alternative methods for three-dimensional (3D) cell culturing. The main modifications concern the cell culture condition and the algorithm that controls the movement of the frames for the nullification of gravity. An incubator was integrated into the inner frame of the RPM allowing precise control over the cell culture environment. Furthermore, several feed-throughs now allow a permanent supply of gas like CO 2. All these modifications substantially improve conditions to culture cells; furthermore, the rewritten software responsible for controlling the movement of the frames enhances the quality of the generated microgravity. Cell culture experiments were carried out with human lymphocytes on the novel RPM model to compare the obtained response to the results gathered on an older well-established RPM as well as to data from space flights. The overall outcome of the tests validates this novel RPM for cell cultivation under simulated microgravity conditions.
-
Moxey, L.; Dias, R. K.; Legaspi, E.
2011-12-01
During the summer of 2011, the Mālama Ke Ahupua`a (to care of our watershed) GEARUP summer program provided 25 under-served and under-represented minority public high school students (Hawaiian, part-Hawaiian, Filipino, Pacific Islanders) from Farrington High School (Kalihi, Honolulu) with a hands-on place-based multidiscipline course located within Manoa Valley (Ahupua`a O Kona) with the objective of engaging participants in scientific environmental investigations while exploring Hawaii's linkages between traditional knowledge, culture and science. The 4-week field program enabled students to collect samples along the perennial Manoa Stream and conduct water quality assessments throughout the Manoa watershed. Students collected science quality data from eight different sampling stations by means of field- and laboratory-based quantitative water quality testing equipment and GPS/GIS technology. While earning Hawaii DOE academic credits, students were able to document changes along the stream as related to pollution and urbanization. While conducting the various scientific investigations, students also participated in cultural fieldtrips and activities that highlighted the linkages between historical sustainable watershed uses by native Hawaiian communities, and their connections with natural earth processes. Additionally, students also participated in environmental service-learning projects that highlight the Hawaiian values of laulima (teamwork), mālama (to care for), and imi `ike (to seek knowledge). By contextualizing and merging hands-on place-based earth science inquiry with native Hawaiian traditional knowledge, students experienced the natural-cultural significance of their ahupua`a (watershed). This highlighted the advantages for promoting environmental literacy and geoscience education to under-served and under-represented minority populations in Hawaii from a rich native Hawaiian cultural framework.
-
Peh, Gary S L; Toh, Kah-Peng; Ang, Heng-Pei; Seah, Xin-Yi; George, Benjamin L; Mehta, Jodhbir S
2013-05-03
Global shortage of donor corneas greatly restricts the numbers of corneal transplantations performed yearly. Limited ex vivo expansion of primary human corneal endothelial cells is possible, and a considerable clinical interest exists for development of tissue-engineered constructs using cultivated corneal endothelial cells. The objective of this study was to investigate the density-dependent growth of human corneal endothelial cells isolated from paired donor corneas and to elucidate an optimal seeding density for their extended expansion in vitro whilst maintaining their unique cellular morphology. Established primary human corneal endothelial cells were propagated to the second passage (P2) before they were utilized for this study. Confluent P2 cells were dissociated and seeded at four seeding densities: 2,500 cells per cm2 ('LOW'); 5,000 cells per cm2 ('MID'); 10,000 cells per cm2 ('HIGH'); and 20,000 cells per cm2 ('HIGH(×2)'), and subsequently analyzed for their propensity to proliferate. They were also subjected to morphometric analyses comparing cell sizes, coefficient of variance, as well as cell circularity when each culture became confluent. At the two lower densities, proliferation rates were higher than cells seeded at higher densities, though not statistically significant. However, corneal endothelial cells seeded at lower densities were significantly larger in size, heterogeneous in shape and less circular (fibroblastic-like), and remained hypertrophic after one month in culture. Comparatively, cells seeded at higher densities were significantly homogeneous, compact and circular at confluence. Potentially, at an optimal seeding density of 10,000 cells per cm2, it is possible to obtain between 10 million to 25 million cells at the third passage. More importantly, these expanded human corneal endothelial cells retained their unique cellular morphology. Our results demonstrated a density dependency in the culture of primary human corneal endothelial
-
Cell cycle progression in irradiated endothelial cells cultured from bovine aorta
International Nuclear Information System (INIS)
Rubin, D.B.; Drab, E.A.; Ward, W.F.; Bauer, K.D.
1988-01-01
Logarithmically growing endothelial cells from bovine aortas were exposed to single doses of 0-10 Gy of 60Co gamma rays, and cell cycle phase distribution and progression were examined by flow cytometry and autoradiography. In some experiments, cells were synchronized in the cell cycle with hydroxyurea (1 mM). Cell number in sham-irradiated control cultures doubled in approximately 24 h. Estimated cycle stage times for control cells were 14.4 h for G1 phase, 7.2 h for S phase, and 2.4 h for G2 + M phase. Irradiated cells demonstrated a reduced distribution at the G1/S phase border at 4 h, and an increased distribution in G2 + M phase at 24 h postirradiation. Autoradiographs of irradiated cells after continuous [3H]thymidine labeling indicated a block in G1 phase or at the G1/S-phase border. The duration of the block was dose dependent (2-3 min/cGy). Progression of the endothelial cells through S phase after removal of the hydroxyurea block also was retarded by irradiation, as demonstrated by increased distribution in early S phase and decreased distribution in late S phase. These results indicate that progression of asynchronous cultured bovine aortic endothelial cells through the DNA synthetic cycle is susceptible to radiation inhibition at specific sites in the cycle, resulting in redistribution and partial synchronization of the population. Thus aortic endothelial cells, diploid cells from a normal tissue, resemble many immortal cell types that have been examined in this regard in vitro
-
Cloning higher plants from aseptically cultured tissues and cells
Krikorian, A. D.
1982-01-01
A review of aseptic culture methods for higher plants is presented, which focuses on the existing problems that limit or prevent the full realization of cloning plants from free cells. It is shown that substantial progress in clonal multiplication has been made with explanted stem tips or lateral buds which can be stimulated to produce numerous precocious axillary branches. These branches can then be separated or subdivided and induced to root in order to yield populations of genetically and phenotypically uniorm plantlets. Similarly, undifferentiated calluses can sometimes be induced to form shoots and/or roots adventitiously. Although the cell culture techniques required to produce somatic embryos are presently rudimentary, steady advances are being made in learning how to stimulate formation of somatic or adventive embryos from totipotent cells grown in suspension cultures. It is concluded that many problems exist in the producing and growing of totipotent or morphogenetically competent cell suspensions, but the potential benefits are great.
-
Culture of human cell lines by a pathogen-inactivated human platelet lysate.
Fazzina, R; Iudicone, P; Mariotti, A; Fioravanti, D; Procoli, A; Cicchetti, E; Scambia, G; Bonanno, G; Pierelli, L
2016-08-01
Alternatives to the use of fetal bovine serum (FBS) have been investigated to ensure xeno-free growth condition. In this study we evaluated the efficacy of human platelet lysate (PL) as a substitute of FBS for the in vitro culture of some human cell lines. PL was obtained by pools of pathogen inactivated human donor platelet (PLT) concentrates. Human leukemia cell lines (KG-1, K562, JURKAT, HL-60) and epithelial tumor cell lines (HeLa and MCF-7) were cultured with either FBS or PL. Changes in cell proliferation, viability, morphology, surface markers and cell cycle were evaluated for each cell line. Functional characteristics were analysed by drug sensitivity test and cytotoxicity assay. Our results demonstrated that PL can support growth and expansion of all cell lines, although the cells cultured in presence of PL experienced a less massive proliferation compared to those grown with FBS. We found a comparable percentage of viable specific marker-expressing cells in both conditions, confirming lineage fidelity in all cultures. Functionality assays showed that cells in both FBS- and PL-supported cultures maintained their normal responsiveness to adriamycin and NK cell-mediated lysis. Our findings indicate that PL is a feasible serum substitute for supporting growth and propagation of haematopoietic and epithelial cell lines with many advantages from a perspective of process standardization, ethicality and product safety.
-
Effects of deprivation of background environmental radiation on cultured human cells
International Nuclear Information System (INIS)
Carbone, M.C.; Pinto, M.; Antonelli, F.; Balata, M.
2010-01-01
In this paper we present results from an experiment aimed at investigating whether living cells are influenced by background ionizing radiation. Parallel human cell cultures were set-up in two separate laboratories and maintained for several months under identical conditions but for a 80 x different level of background ionizing radiation. Periodically, the cell cultures were monitored for the onset of divergences in biochemical behavior, using two distinct cellular biology assays, namely micronuclei induction and activity of enzymes implicated in the management of oxidative stress. To reveal any subtle modifications, responses were also amplified by subjecting cell cultures to acute stress induced by exposure to moderately high doses of ionizing radiation. Compared to reference radiation background conditions, cultures maintained in a reduced background radiation environment handled the consequences of acute stress with diminished efficacy.
-
Karyometric observations of WISH cell cultures irradiated with 3 GHz microwaves
Energy Technology Data Exchange (ETDEWEB)
Szmigielski, S.; Luczak, M.; Wiranowska, M.
1975-01-01
WISH cell cultures 24 hours after passage were irradiated with 3 GHz microwaves (10 cm) at far field conditions in free space (anechoic chamber) for 30 minutes, at field power density 5 or 20 mW/cm/sup 2/. Within 1, 24, and 48 hours of the exposure to microwave fields the volumes of nuclei and nucleoli were measured with the use of a micrometer, and logvolumes and nucleo-nucleolar ratios were calculated. Under the applied irradiation conditions the culture medium temperature did not exceed 37/sup 0/C. In cultures irradiated at field power density 20 mW/cm/sup 2/ increased number of cells with small nuclei and enlarged nucleoli was noted within 1 hour of the exposure. Within 24 and 48 hours after irradiation the nucleolar volume showed a slight decrease, whereas the nuclear volume increased. In cultures irradiated at field power density 5 mW/cm/sup 2/ increased numbers of cells with enlarged nuclei and nucleoli were found. Analysis of the distribution curves of nuclear and nucleolar volumes suggests that non-thermal power densities of microwaves stimulate the metabolism of cell cultures. However, at higher power densities (20 mW/cm/sup 2/) the stimulation phase is preceded by a period of reduced viability of cell cultures.
-
Tailor-made three-dimensional hybrid scaffolds for cell cultures
Energy Technology Data Exchange (ETDEWEB)
Psycharakis, Stylianos; Melissinaki, Vasileia; Giakoumaki, Anastasia; Ranella, Anthi [Institute of Electronic Structure and Laser, Foundation for Research and Technology Hellas, PO Box 1527, 711 10 Heraklion, Crete (Greece); Tosca, Androniki, E-mail: ranthi@iesl.forth.gr [Department of Medicine, University of Crete, 710 03 Heraklion, Crete (Greece)
2011-08-15
The construction of the ideal three-dimensional scaffold for cell culture is one of the most intriguing topics in tissue engineering. It has been shown that cells can be cultured on most organic biomimetic materials, which now are losing popularity in favour of novel, hybrid systems. In this study, a series of photosensitive sol-gel hybrid materials, based on silicon-zirconium and silicon-titanium oxides, have been investigated for their suitability in three-dimensional scaffold fabrication. These materials can be structured by two-photon polymerization, a laser-based technique allowing the fabrication of micrometre-size structures with submicron resolution. The work presented here examined the effect of the organic/inorganic composition of the materials on cell behaviour and the establishment of a 'cell-culture friendly' environment. This is vital for cell adhesion, growth and differentiation, as the organic part of the material provides the soft matrix for cell growth, whereas the inorganic component gives the mechanical stability and rigidity of the three-dimensional structures. In addition, the use of femtosecond laser structuring permits the fabrication of a wide range of mechanically stable scaffolds of different sizes and shapes to be tested in terms of cell viability, proliferation and orientation.
-
Directory of Open Access Journals (Sweden)
Corneo Gianmarco
2007-07-01
Full Text Available Abstract Background Tumor cell contamination of the apheresis in multiple myeloma is likely to affect disease-free and overall survival after autografting. Objective To purge myeloma aphereses from tumor contaminants with a novel culture-based purging method. Methods We cultured myeloma-positive CD34+ PB samples in conditions that retained multipotency of hematopoietic stem cells, but were unfavourable to survival of plasma cells. Moreover, we exploited the resistance of myeloma plasma cells to retroviral transduction by targeting the hematopoietic CD34+ cell population with a retroviral vector carrying a selectable marker (the truncated form of the human receptor for nerve growth factor, ΔNGFR. We performed therefore a further myeloma purging step by selecting the transduced cells at the end of the culture. Results Overall recovery of CD34+ cells after culture was 128.5%; ΔNGFR transduction rate was 28.8% for CD34+ cells and 0% for CD138-selected primary myeloma cells, respectively. Recovery of CD34+ cells after ΔNGFR selection was 22.3%. By patient-specific Ig-gene rearrangements, we assessed a decrease of 0.7–1.4 logs in tumor load after the CD34+ cell selection, and up to 2.3 logs after culture and ΔNGFR selection. Conclusion We conclude that ex-vivo culture and retroviral-mediated transduction of myeloma leukaphereses provide an efficient tumor cell purging.
-
Tan, Kah Yong; Teo, Kim Leng; Lim, Jessica F Y; Chen, Allen K L; Choolani, Mahesh; Reuveny, Shaul; Chan, Jerry; Oh, Steve Kw
2015-08-01
Mesenchymal stromal cells (MSCs) are being investigated as potential cell therapies for many different indications. Current methods of production rely on traditional monolayer culture on tissue-culture plastic, usually with the use of serum-supplemented growth media. However, the monolayer culturing system has scale-up limitations and may not meet the projected hundreds of billions to trillions batches of cells needed for therapy. Furthermore, serum-free medium offers several advantages over serum-supplemented medium, which may have supply and contaminant issues, leading to many serum-free medium formulations being developed. We cultured seven MSC lines in six different serum-free media and compared their growth between monolayer and microcarrier culture. We show that (i) expansion levels of MSCs in serum-free monolayer cultures may not correlate with expansion in serum-containing media; (ii) optimal culture conditions (serum-free media for monolayer or microcarrier culture) differ for each cell line; (iii) growth in static microcarrier culture does not correlate with growth in stirred spinner culture; (iv) and that early cell attachment and spreading onto microcarriers does not necessarily predict efficiency of cell expansion in agitated microcarrier culture. Current serum-free media developed for monolayer cultures of MSCs may not support MSC proliferation in microcarrier cultures. Further optimization in medium composition will be required for microcarrier suspension culture for each cell line. Copyright © 2015 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.
-
Schwarz, R. P.; Goodwin, T. J.; Wolf, D. A.
1992-01-01
High-density, three-dimensional cell cultures are difficult to grow in vitro. The rotating-wall vessel (RWV) described here has cultured BHK-21 cells to a density of 1.1 X 10(7) cells/ml. Cells on microcarriers were observed to grow with enhanced bridging in this batch culture system. The RWV is a horizontally rotated tissue culture vessel with silicon membrane oxygenation. This design results in a low-turbulence, low-shear cell culture environment with abundant oxygenation. The RWV has the potential to culture a wide variety of normal and neoplastic cells.
-
Long-Term Oocyte-Like Cell Development in Cultures Derived from Neonatal Marmoset Monkey Ovary
Directory of Open Access Journals (Sweden)
Bentolhoda Fereydouni
2016-01-01
Full Text Available We use the common marmoset monkey (Callithrix jacchus as a preclinical nonhuman primate model to study reproductive and stem cell biology. The neonatal marmoset monkey ovary contains numerous primitive premeiotic germ cells (oogonia expressing pluripotent stem cell markers including OCT4A (POU5F1. This is a peculiarity compared to neonatal human and rodent ovaries. Here, we aimed at culturing marmoset oogonia from neonatal ovaries. We established a culture system being stable for more than 20 passages and 5 months. Importantly, comparative transcriptome analysis of the cultured cells with neonatal ovary, embryonic stem cells, and fibroblasts revealed a lack of germ cell and pluripotency genes indicating the complete loss of oogonia upon initiation of the culture. From passage 4 onwards, however, the cultured cells produced large spherical, free-floating cells resembling oocyte-like cells (OLCs. OLCs strongly expressed several germ cell genes and may derive from the ovarian surface epithelium. In summary, our novel primate ovarian cell culture initially lacked detectable germ cells but then produced OLCs over a long period of time. This culture system may allow a deeper analysis of early phases of female primate germ cell development and—after significant refinement—possibly also the production of monkey oocytes.
-
Kamperman, Tom; Henke, Sieger; Visser, Claas Willem; Karperien, Marcel; Leijten, Jeroen
2017-01-01
Single-cell-laden microgels support physiological 3D culture conditions while enabling straightforward handling and high-resolution readouts of individual cells. However, their widespread adoption for long-term cultures is limited by cell escape. In this work, it is demonstrated that cell escape is
-
Neuron-specific enolase is a useful maker of neuroendocrine origin in pheochromocytoma cell culture
International Nuclear Information System (INIS)
Abelin, N.; Dahia, P.L.M.; Martin, R.; Kato, S.; Toledo, S.P.A.
1994-01-01
Neuron-specific enolase (NSE) has been used as a marker for neuroendocrine tumors either in immunocytochemical studies or in serum measurements. In this paper NSE levels were determined in cultured pheochromocytoma cells to test whether it is also a useful marker in cell culture of tumors derived from neuroendocrine system. Cultured pheochromocytoma cells came from a primary explant and were grown in RPMI supplemented with 20% fetal calf serum, 100 μg/mL ampicillin and 100 μ/mL streptomycin. NSE was measured in culture medium and cell homogenates. Samples from different pheochromocytoma cultures were analyzed and compared to normal cultured fibroblast cells derived from human skin. NSE was measured by a commercially available radioimmunoassay kit. NSE levels were higher in cell homogenates as compared to those in culture medium, reaching levels as high as 6-fold in the former in TE cell line (26.46 ng/mL and 4.39 ng/mL, respectively). Serial measurements in culture medium from TE cell line evidenced decreasing values in subsequential subcultures (from 9.24 ng/mL during primary explant to 1.7 ng/mL in the tenth subculture). In cultured normal fibroblasts, NSE levels in cultured media were definitely lower than those obtained from pheochromocytoma cultures. These preliminary data suggest that NSE may be a useful marker of neuroendocrine derived tumors, such as pheochromocytoma, in culture. Thus, the simplicity and availability of NSE radioimmunoassay provides an alternative to catecholamine measurement to better characterize pheochromocytoma cell lines in culture, with the advantage of faster result at lower costs. (author). 18 refs, 2 tabs
-
Bryson, Dennis
2009-01-01
The field of personality and culture was given a significant impetus during the 1930s with the establishment of the Advisory Committee on Personality and Culture (1930-1934) by the Social Science Research Council. This committee provided an early formulation of personality and culture that emphasized the interdisciplinary focus on the processes of personality formation within small-scale social settings. The committee's formulation also coupled personality and culture with a liberal social engineering approach geared toward cultural reconstruction. Major social scientists and clinicians were involved in the activities of the committee, including Edward Sapir, W. I. Thomas, E. W. Burgess, E. A. Bott, Robert S. Woodworth, Harry Stack Sullivan, C. M. Hincks, and Adolf Meyer.
-
Students′ Perception of Organization Culture at a Faculty of Science and Technology
Directory of Open Access Journals (Sweden)
Maria Ujhelyi
2017-04-01
Full Text Available The current study uses an adapted version of Cameron and Quinn’s OCAI questionnaire to test the organisational culture of the Faculty of Science and Technology at the University of Debrecen, Hungary, as it is perceived by its students, and also to discover what type of organisational culture the same students think would be ideal for them. An additional objective of this paper is to identify possible gaps between the perceived and the ideal cultures expressed by the students. Our sample includes 128 questionnaires completed by bachelor students from 6 different majors at the faculty. According to our results, the respondents perceive to a significant degree that the faculty’s organisational culture is at an average level of clan, market and hierarchy cultures, while it also exhibits a relatively low level of the adhocracy culture. Their ideal faculty culture would be one with average adhocracy, average hierarchy, high clan and low market features. Significant gaps are identified between the perceived and ideal cultures in all the four types: students would prefer an increase in clan and adhocracy cultures, and a decrease in the other two cultures.
-
Do rice suspension-cultured cells treated with abscisic acid mimic developing seeds?
Matsuno, Koya; Fujimura, Tatsuhito
2015-08-01
Starch synthesis is activated in the endosperm during seed development and also in rice suspension cells cultured with abscisic acid. In the anticipation that the mechanisms of starch synthesis are similar between the endosperm and the suspension cells cultured with abscisic acid, expression of genes involved in starch synthesis was evaluated in the suspension cells after abscisic acid treatment. However, it was found that the regulatory mechanism of starch synthesis in the suspension cells cultured with abscisic acid was different from that in developing seeds. Expression analyses of genes involved in oil bodies, which accumulate in the embryo and aleurone layer, and seed storage proteins, which accumulate mainly in the endosperm, showed that the former were activated in the suspension cells cultured with abscisic acid, but the latter were not. Master regulators for embryogenesis, OsVP1 (homologue of AtABI3) and OsLFL1 (homologue of AtFUS3 or AtLFL2), were expressed in the suspension cells at levels comparable to those in the embryo. From these results, it is suggested that interactions between regulators and abscisic acid control the synthesis of phytic acid and oil bodies in the cultured cells and embryo. We suggest that the system of suspension cells cultured with abscisic acid helps to reveal the mechanisms of phytic acid and oil body synthesis in embryo.
-
Effect of anabolics on bovine granulosa-luteal cell primary cultures.
Directory of Open Access Journals (Sweden)
Bartolomeo Biolatti
2007-10-01
Full Text Available Granulosa cell tumours are observed with increased frequency among calves slaughtered in Northern Italy. The use of illegal anabolics in breeding was taken into account as a cause of this pathology. An in vitro approach was used to detect the possible alterations of cell proliferation induced by anabolics on primary cultures of bovine granulosa-luteal cells. Cultures were treated with different concentrations of substances illegally used in cattle (17beta-estradiol, clenbuterol and boldione. Cytotoxicity was determined by means of MTT test, to exclude toxic effects induced by anabolics and to determine the highest concentration to be tested. Morphological changes were evaluated by means of routine cytology, while PCNA expression was quantified in order to estimate cell proliferation. Cytotoxic effects were revealed at the highest concentrations. The only stimulating effect on cell proliferation was detected in boldione treated cultures: after 48 h treated cells, compared to controls, showed a doubled expression of PCNA. In clenbuterol and 17beta-estradiol treated cells PCNA expression was similar to controls or even decreased. As the data suggest an alteration in cell proliferation, boldione could have a role in the early stage of pathogenesis of granulosa cell tumour in cattle.
-
Park, Ji Eun; Park, Min Hee; Kim, Min Seong; Park, Yeo Reum; Yun, Jung Im; Cheong, Hee Tae; Kim, Minseok; Choi, Jung Hoon; Lee, Eunsong; Lee, Seung Tae
2017-12-01
Generally, self-renewal of spermatogonial stem cells (SSCs) is maintained in vivo in a three-dimensional (3D) microenvironment consisting of the seminiferous tubule basement membrane, indicating the importance of the 3D microenvironment for in vitro culture of SSCs. Here, we report a 3D culture microenvironment that effectively maintains porcine SSC self-renewal during culture. Porcine SSCs were cultured in an agarose-based 3D hydrogel and in 2D culture plates either with or without feeder cells. Subsequently, the effects of 3D culture on the maintenance of undifferentiated SSCs were identified by analyzing cell colony formation and morphology, AP activity, and transcriptional and translational regulation of self-renewal-related genes and the effects on proliferation by analyzing cell viability and single cell-derived colony number. The 3D culture microenvironment constructed using a 0.2% (w/v) agarose-based 3D hydrogel showed the strongest maintenance of porcine SSC self-renewal and induced significant improvements in proliferation compared with 2D culture microenvironments. These results demonstrate that self-renewal of porcine SSCs can be maintained more effectively in a 3D than in a 2D culture microenvironment. Moreover, this will play a significant role in developing novel culture systems for SSCs derived from diverse species in the future, which will contribute to SSC-related research. © 2017 International Federation for Cell Biology.
-
Establishment of Cancer Stem Cell Cultures from Human Conventional Osteosarcoma.
Palmini, Gaia; Zonefrati, Roberto; Mavilia, Carmelo; Aldinucci, Alessandra; Luzi, Ettore; Marini, Francesca; Franchi, Alessandro; Capanna, Rodolfo; Tanini, Annalisa; Brandi, Maria Luisa
2016-10-14
The current improvements in therapy against osteosarcoma (OS) have prolonged the lives of cancer patients, but the survival rate of five years remains poor when metastasis has occurred. The Cancer Stem Cell (CSC) theory holds that there is a subset of tumor cells within the tumor that have stem-like characteristics, including the capacity to maintain the tumor and to resist multidrug chemotherapy. Therefore, a better understanding of OS biology and pathogenesis is needed in order to advance the development of targeted therapies to eradicate this particular subset and to reduce morbidity and mortality among patients. Isolating CSCs, establishing cell cultures of CSCs, and studying their biology are important steps to improving our understanding of OS biology and pathogenesis. The establishment of human-derived OS-CSCs from biopsies of OS has been made possible using several methods, including the capacity to create 3-dimensional stem cell cultures under nonadherent conditions. Under these conditions, CSCs are able to create spherical floating colonies formed by daughter stem cells; these colonies are termed "cellular spheres". Here, we describe a method to establish CSC cultures from primary cell cultures of conventional OS obtained from OS biopsies. We clearly describe the several passages required to isolate and characterize CSCs.
-
Yamaguchi, Y; Kluge, N; Ostertag, W; Furusawa, M
1981-01-01
Cell cultures of 7,12-dimethylbenz[a]anthracene-induced rat erythroleukemia can be stimulated to synthesize hemoglobin when cultured in hypertonic media. During hypertonic treatment the intracellular osmotic conditions immediately readjust to those of the extracellular medium. None of the Friend virus-induced mouse erythroleukemia cell lines was inducible for differentiation with the same hypertonic culture conditions used for rat cells. Earliest commitment to erythroid terminal differentiati...
-
International Nuclear Information System (INIS)
Oberley, T.D.; Lauchner, L.J.; Pugh, T.D.; Gonzalez, A.; Goldfarb, S.; Li, S.A.; Li, J.J.
1989-01-01
It has long been recognized that the renal proximal tubular epithelium of the hamster is a bona fide estrogen target tissue. The effect of estrogens on the growth of proximal tubule cell explants and dissociated single cells derived from these explant outgrowths has been studied in culture. Renal tubular cells were grown on a PF-HR-9 basement membrane under serum-free chemically defined culture conditions. At 7-14 days in culture, cell number was enhanced 3-fold in the presence of either 17β-estradiol or diethylstilbestrol. A similar 3-fold increase in cell number was also seen at 1 nM 17β-estradiol in subcultured dissociated single tubular cells derived from hamster renal tubular explant outgrowths at 21 days in culture. Concomitant exposure of tamoxifen at 3-fold molar excess in culture completely abolished the increase in cell number seen with 17β-estradiol. The proliferation effect of estrogens on proximal tubular cell growth appears to be species specific since 17β-estradiol did not alter the growth of either rat or guinea pig proximal tubules in culture. In addition, at 7-10 days in culture in the presence of 17β-estradiol, [ 3 H]thymidine labeling of hamster tubular cells was enhanced 3-fold. These results clearly indicate that estrogens can directly induce primary epithelial cell proliferation at physiologic concentrations and provide strong additional evidence for an important hormonal role in the neoplastic transformation of the hamster kidney
-
Transfection in perfused microfluidic cell culture devices: A case study.
Raimes, William; Rubi, Mathieu; Super, Alexandre; Marques, Marco P C; Veraitch, Farlan; Szita, Nicolas
2017-08-01
Automated microfluidic devices are a promising route towards a point-of-care autologous cell therapy. The initial steps of induced pluripotent stem cell (iPSC) derivation involve transfection and long term cell culture. Integration of these steps would help reduce the cost and footprint of micro-scale devices with applications in cell reprogramming or gene correction. Current examples of transfection integration focus on maximising efficiency rather than viable long-term culture. Here we look for whole process compatibility by integrating automated transfection with a perfused microfluidic device designed for homogeneous culture conditions. The injection process was characterised using fluorescein to establish a LabVIEW-based routine for user-defined automation. Proof-of-concept is demonstrated by chemically transfecting a GFP plasmid into mouse embryonic stem cells (mESCs). Cells transfected in the device showed an improvement in efficiency (34%, n = 3) compared with standard protocols (17.2%, n = 3). This represents a first step towards microfluidic processing systems for cell reprogramming or gene therapy.
-
International Nuclear Information System (INIS)
Kim, Jae Sung; Lee, Young Keun; Chung, Byung Yeoup; Lee, Young Bok; Hwang Hye Yeon
2004-10-01
The cell lines 679, 679-29 and 622-46 of L. erythrorhizon could be selected on LS agar medium for the production shikonin in cell suspension culture. The shikonin was increased moderately in suspension culture of cell line 622-46 in LS liquid medium containing BA 2 mg·L -1 and IAA 0.2 mg·L -1 in the dark, and was increased by adding 1 μM Cu 2+ and 100 μM methyl jasmonate The accumulation of shikonin in the liquid medium was increased significantly by 2 Gy irradiation to callus of cell line 622-46 and culture in LS liquid medium containing BA 2 mg·L -1 and IAA 0.2 mg·L -1 in the dark and shikonin in cell debris was higher by 16 Gy irradiation. The activity of p-hydroxybenzoate geranyltransferase was increased by irradiation of 2 Gy and 16 Gy of γ radiation. Seedling hypocotyles of L. erythrorhizon were infected with Agrogacterium rhizogenes strain 15834 harboring a binary vector with an intron bearing the GUS (β-glucuronidase) gene driven by cauliflower mosaic virus (CaMV) 35S promotor as well as the HPT (hygromycin phosphotransferase) gene as the selection marker. Hairy roots isolated were hygromycin resistant and had integrated GUS gene in DNA. The root tip grown on M-9 medium showed normal pigment production pattern in border cells and root hairs
-
Directory of Open Access Journals (Sweden)
William Lee
2008-12-01
Full Text Available Astrocytes play an important role in cell-cell signaling in the mammalian central nervous system. The ability of astrocytes to communicate with surrounding cells through gap-junctional coupling or signaling via the release of transmitters makes characterization of these cells difficult in vitro and even more so in vivo. To simplify the complexity of common in vitro systems, introduced by intercellular communication between astrocytes, we developed a novel cell culturing method, in which purified rat visual cortical astrocytes were grown in spatially defined cell-adhesion wells which we termed micropits. We showed that astrocytes cultured in micropit regions were viable and exhibited similar characteristics of Ca2+ dynamics and astrocytic marker expression to those of cells cultured in non-micropit regions. Examination of intracellular Ca2+ oscillations in solitary astrocytes cultured in micropits revealed less variable oscillations than those of non-micropit grouped astrocytes, which were in contact with their neighbors. Solitary cells in micropit regions can undergo ATP-mediated astrocyte-microglia signaling, demonstrating that this culturing method can also be used to investigate glial-glial interactions in a spatially well-defined microenvironment.
-
Aromatase inhibitor (anastrozole) affects growth of endometrioma cells in culture.
Badawy, Shawky Z A; Brown, Shereene; Kaufman, Lydia; Wojtowycz, Martha A
2015-05-01
To study the effects of aromatase inhibitor (anastrozole) on the growth and estradiol secretion of endometrioma cells in culture. Endometrioma cells are grown in vitro until maximum growth before used in this study. This was done in the research laboratory for tissue culture, in an academic hospital. Testosterone at a concentration of 10 μg/mL was added as a substrate for the intracellular aromatase. In addition, aromatase inhibitor was added at a concentration of 200 and 300 μg/mL. The effect on cell growth and estradiol secretion is evaluated using Student's t-test. The use of testosterone increased estradiol secretion by endometrioma cells in culture. The use of aromatase inhibitor significantly inhibited the growth of endometrioma cells, and estradiol secretion. Aromatase inhibitor (anastrozole) may be an effective treatment for endometriosis due to inhibition of cellular aromatase. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.
-
Keyvanara, Mahmoud; Yarmohammadian, Mohammad Hossein; Soltani, Batoul
2014-01-01
Cultural intelligence and social compatibility are two acquired processes that their education and reinforcement between dormitory's students who have inter cultural interactions with each other can conclude with results that tension diminution, inter cultural contrast and conflict, social divisions and consequently healthy and peaceful relationships and governance and finally mental peace, and health are of its most important. Hence, the research has been occurring in order to the determination of cultural intelligence relationship with the social compatibility of Isfahan University of Medical Sciences dormitories resident students in 2012. The research method is descriptive-correlation, and its population is composed of all Isfahan University of Medical Sciences dormitories resident students in 2012 that were totally 2500 persons. The two steps sampling method have been used, group sampling and random sampling has been occurring at first and second steps and totally 447 persons were selected. Research data were collected via Earley and Ang cultural intelligence questionnaire with 0.76 Cronbach's alpha Coefficient and California social compatibility standard questionnaire with higher than 0.70 Cronbach's alpha factor. Questionnaire data have been analyzed with the SPSS software and results have been presented in the shape of descriptions and statistics. Results showed that there is a direct significant relationship (P intelligence and the social adjustment in students living in Isfahan University of Medical Sciences dormitories and also there is a direct significant relationship in the level of (P intelligence; however, there is no significant relationship between cognitive and behavioral dimensions of cultural intelligence and social adjustment (P > 0.05). Cultural intelligence and cognitive and motivational addition in dimensions of students living in Isfahan University of Medical Sciences dormitories increase their social integration, therefore, cultural
-
Automation of 3D cell culture using chemically defined hydrogels.
Rimann, Markus; Angres, Brigitte; Patocchi-Tenzer, Isabel; Braum, Susanne; Graf-Hausner, Ursula
2014-04-01
Drug development relies on high-throughput screening involving cell-based assays. Most of the assays are still based on cells grown in monolayer rather than in three-dimensional (3D) formats, although cells behave more in vivo-like in 3D. To exemplify the adoption of 3D techniques in drug development, this project investigated the automation of a hydrogel-based 3D cell culture system using a liquid-handling robot. The hydrogel technology used offers high flexibility of gel design due to a modular composition of a polymer network and bioactive components. The cell inert degradation of the gel at the end of the culture period guaranteed the harmless isolation of live cells for further downstream processing. Human colon carcinoma cells HCT-116 were encapsulated and grown in these dextran-based hydrogels, thereby forming 3D multicellular spheroids. Viability and DNA content of the cells were shown to be similar in automated and manually produced hydrogels. Furthermore, cell treatment with toxic Taxol concentrations (100 nM) had the same effect on HCT-116 cell viability in manually and automated hydrogel preparations. Finally, a fully automated dose-response curve with the reference compound Taxol showed the potential of this hydrogel-based 3D cell culture system in advanced drug development.
-
An efficient 3D cell culture method on biomimetic nanostructured grids.
Directory of Open Access Journals (Sweden)
Maria Wolun-Cholewa
Full Text Available Current techniques of in vitro cell cultures are able to mimic the in vivo environment only to a limited extent, as they enable cells to grow only in two dimensions. Therefore cell culture approaches should rely on scaffolds that provide support comparable to the extracellular matrix. Here we demonstrate the advantages of novel nanostructured three-dimensional grids fabricated using electro-spinning technique, as scaffolds for cultures of neoplastic cells. The results of the study show that the fibers allow for a dynamic growth of HeLa cells, which form multi-layer structures of symmetrical and spherical character. This indicates that the applied scaffolds are nontoxic and allow proper flow of oxygen, nutrients, and growth factors. In addition, grids have been proven to be useful in in situ examination of cells ultrastructure.
-
Kihara, Takanori; Kashitani, Kosuke; Miyake, Jun
2017-07-14
Cell proliferation is a key characteristic of eukaryotic cells. During cell proliferation, cells interact with each other. In this study, we developed a cellular automata model to estimate cell-cell interactions using experimentally obtained images of cultured cells. We used four types of cells; HeLa cells, human osteosarcoma (HOS) cells, rat mesenchymal stem cells (MSCs), and rat smooth muscle A7r5 cells. These cells were cultured and stained daily. The obtained cell images were binarized and clipped into squares containing about 10 4 cells. These cells showed characteristic cell proliferation patterns. The growth curves of these cells were generated from the cell proliferation images and we determined the doubling time of these cells from the growth curves. We developed a simple cellular automata system with an easily accessible graphical user interface. This system has five variable parameters, namely, initial cell number, doubling time, motility, cell-cell adhesion, and cell-cell contact inhibition (of proliferation). Within these parameters, we obtained initial cell numbers and doubling times experimentally. We set the motility at a constant value because the effect of the parameter for our simulation was restricted. Therefore, we simulated cell proliferation behavior with cell-cell adhesion and cell-cell contact inhibition as variables. By comparing growth curves and proliferation cell images, we succeeded in determining the cell-cell interaction properties of each cell. Simulated HeLa and HOS cells exhibited low cell-cell adhesion and weak cell-cell contact inhibition. Simulated MSCs exhibited high cell-cell adhesion and positive cell-cell contact inhibition. Simulated A7r5 cells exhibited low cell-cell adhesion and strong cell-cell contact inhibition. These simulated results correlated with the experimental growth curves and proliferation images. Our simulation approach is an easy method for evaluating the cell-cell interaction properties of cells.
-
de Almeida-Leite, Camila Megale; Arantes, Rosa Maria Esteves
2010-12-15
Central nervous system glial cells as astrocytes and microglia have been investigated in vitro and many intracellular pathways have been clarified upon various stimuli. Peripheral glial cells, however, are not as deeply investigated in vitro despite its importance role in inflammatory and neurodegenerative diseases. Based on our previous experience of culturing neuronal cells, our objective was to standardize and morphologically characterize a primary culture of mouse superior cervical ganglion glial cells in order to obtain a useful tool to study peripheral glial cell biology. Superior cervical ganglia from neonatal C57BL6 mice were enzymatically and mechanically dissociated and cells were plated on diluted Matrigel coated wells in a final concentration of 10,000cells/well. Five to 8 days post plating, glial cell cultures were fixed for morphological and immunocytochemical characterization. Glial cells showed a flat and irregular shape, two or three long cytoplasm processes, and round, oval or long shaped nuclei, with regular outline. Cell proliferation and mitosis were detected both qualitative and quantitatively. Glial cells were able to maintain their phenotype in our culture model including immunoreactivity against glial cell marker GFAP. This is the first description of immunocytochemical characterization of mouse sympathetic cervical ganglion glial cells in primary culture. This work discusses the uses and limitations of our model as a tool to study many aspects of peripheral glial cell biology. Copyright © 2010 Elsevier B.V. All rights reserved.
-
Palethorpe, Helen M; Leach, Damien A; Need, Eleanor F; Drew, Paul A; Smith, Eric
2018-04-10
Fibroblasts express androgen receptor (AR) in the normal prostate and during prostate cancer development. We have reported that loss of AR expression in prostate cancer-associated fibroblasts is a poor prognostic indicator. Here we report outcomes of direct and indirect co-cultures of immortalised AR-positive (PShTert-AR) or AR-negative (PShTert) myofibroblasts with prostate cancer cells. In the initial co-cultures the AR-negative PC3 cell line was used so AR expression and signalling were restricted to the myofibroblasts. In both direct and indirect co-culture with PShTert-AR myofibroblasts, paracrine signalling to the PC3 cells slowed proliferation and induced apoptosis. In contrast, PC3 cells proliferated with PShTert myofibroblasts irrespective of the co-culture method. In direct co-culture PC3 cells induced apoptosis in and destroyed PShTerts by direct signalling. Similar results were seen in direct co-cultures with AR-negative DU145 and AR-positive LNCaP and C4-2B prostate cancer cell lines. The AR ligand 5α-dihydrotestosterone (DHT) inhibited the proliferation of the PShTert-AR myofibroblasts, thereby reducing the extent of their inhibitory effect on cancer cell growth. These results suggest loss of stromal AR would favour prostate cancer cell growth in vivo , providing an explanation for the clinical observation that reduced stromal AR is associated with a poorer outcome.
-
Multistage carcinogenesis in cell culture.
Rubin, H
2001-01-01
Rodent fibroblasts explanted from embryos to culture undergo a period of declining growth rate in serial passages leading to crisis, followed by the appearance of variants which can multiply indefinitely. If the "immortal" cell line was established by low density passage, i.e., 3T3 cells, it has a low saturation density and is non-tumorigenic. If it was established by high density passage, it has a high saturation density and is tumorigenic. The establishment of cells goes through successive stages, including increased capacity to multiply in low serum concentration, growth to high saturation density, growth in suspension, assisted tumour formation in susceptible hosts and unassisted tumour formation. Chromosome aberrations and aneuploidy occur long before the capacity to produce tumours appears. Contrary to conventional belief, human fibroblast populations also undergo a continuous loss of capacity to multiply from the time of explantation, with only the longest surviving clone reaching the Hayflick limit. Neoplastic transformation of rodent cells is strongly favoured by maintaining them in a quiescent state at confluence for prolonged periods, which results in genetic damage to the cells. It also produces a large variety of chromosomal aberrations in human cells and extends their replicative lifespan. Individual clones are more susceptible to spontaneous transformation than their heterogeneous parental cultures. The implications of these results for tumour development in vivo are that oncogenic genetic changes may be common under stressful conditions which restrict replication, and that such changes are maximized when a rogue clone reaches a critical size that reduces stabilizing interactions with neighbouring clones. An alternative explanation, described in the Addendum, which we retrospectively favor is that the easily transformed clones are a minority in the uncloned parental population. The reason they transform before the parental population is that when
-
Saha, Krishanu; Mei, Ying; Reisterer, Colin M; Pyzocha, Neena Kenton; Yang, Jing; Muffat, Julien; Davies, Martyn C; Alexander, Morgan R; Langer, Robert; Anderson, Daniel G; Jaenisch, Rudolf
2011-11-15
The current gold standard for the culture of human pluripotent stem cells requires the use of a feeder layer of cells. Here, we develop a spatially defined culture system based on UV/ozone radiation modification of typical cell culture plastics to define a favorable surface environment for human pluripotent stem cell culture. Chemical and geometrical optimization of the surfaces enables control of early cell aggregation from fully dissociated cells, as predicted from a numerical model of cell migration, and results in significant increases in cell growth of undifferentiated cells. These chemically defined xeno-free substrates generate more than three times the number of cells than feeder-containing substrates per surface area. Further, reprogramming and typical gene-targeting protocols can be readily performed on these engineered surfaces. These substrates provide an attractive cell culture platform for the production of clinically relevant factor-free reprogrammed cells from patient tissue samples and facilitate the definition of standardized scale-up friendly methods for disease modeling and cell therapeutic applications.
-
Aragonite precipitation by "proto-polyps" in coral cell cultures.
Directory of Open Access Journals (Sweden)
Tali Mass
Full Text Available The mechanisms of coral calcification at the molecular, cellular and tissue levels are poorly understood. In this study, we examine calcium carbonate precipitation using novel coral tissue cultures that aggregate to form "proto-polyps". Our goal is to establish an experimental system in which calcification is facilitated at the cellular level, while simultaneously allowing in vitro manipulations of the calcifying fluid. This novel coral culturing technique enables us to study the mechanisms of biomineralization and their implications for geochemical proxies. Viable cell cultures of the hermatypic, zooxanthellate coral, Stylophora pistillata, have been maintained for 6 to 8 weeks. Using an enriched seawater medium with aragonite saturation state similar to open ocean surface waters (Ω(arag~4, the primary cell cultures assemble into "proto-polyps" which form an extracellular organic matrix (ECM and precipitate aragonite crystals. These extracellular aragonite crystals, about 10 µm in length, are formed on the external face of the proto-polyps and are identified by their distinctive elongated crystallography and X-ray diffraction pattern. The precipitation of aragonite is independent of photosynthesis by the zooxanthellae, and does not occur in control experiments lacking coral cells or when the coral cells are poisoned with sodium azide. Our results demonstrate that proto-polyps, aggregated from primary coral tissue culture, function (from a biomineralization perspective similarly to whole corals. This approach provides a novel tool for investigating the biophysical mechanism of calcification in these organisms.
-
International Nuclear Information System (INIS)
Haubner, Frank; Leyh, Michaela; Ohmann, Elisabeth; Pohl, Fabian; Prantl, Lukas; Gassner, Holger G
2013-01-01
The inflammatory response clinically observed after radiation has been described to correlate with elevated expression of cytokines and adhesion molecules by endothelial cells. Therapeutic compensation for this microvascular compromise could be an important approach in the treatment of irradiated wounds. Clinical reports describe the potential of adipose-derived stem cells to enhance wound healing, but the underlying cellular mechanisms remain largely unclear. Human dermal microvascular endothelial cells (HDMEC) and human adipose-derived stem cells (ASC) were cultured in a co-culture setting and irradiated with sequential doses of 2 to 12 Gy. Cell count was determined 48 h after radiation using a semi-automated cell counting system. Levels of interleukin-6 (IL-6), basic fibroblast growth factor (FGF), intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) were determined in the supernatants using enzyme-linked immunosorbent assay (ELISA). Irradiated HDMEC and ASC as well as non-irradiated co-cultures, HDMEC or ASC respectively were used as controls. Cell count was significantly reduced in irradiated co-cultures of HDMEC and ASC compared to non-irradiated controls. Levels of IL-6, FGF, ICAM-1 and VCAM-1 in the supernatants of the co-cultures were significantly less affected by external radiation in comparison to HDMEC. The increased expression of cytokines and adhesion molecules by HDMEC after external radiation is mitigated in the co-culture setting with ASC. These in vitro changes seem to support the clinical observation that ASC may have a stabilizing effect when injected into irradiated wounds
-
Bioethics of Universal Knowledge: How Space Science is Transforming Global Culture
Perkins, Kala
A new universal culture is championing the human race; never before has immersion in the cosmological environment been so clearly presented nor invited as revolutionary a sense of participatory identity to the human race. We are delving into the awareness of a complex relatedness with the expanse of spatial architectures and life that astrophysics and cosmology are revealing. History is marked by waves of interest and inquiry into the possibilities of the existence of other worlds. Since the Renaissance, building of telescopes has been pursued in their quest; now Kepler and other space missions are leading us into direct apprehension of these worlds, scattered across the cosmological landscape. This affords a unique repertoire of dimensionalities in which to re-construe our global cultural evolution and identity. Spatial education, with related social science and humanities, are facilitating the actualization of a universal culture, redefining the collective global heritage, with infinity as our home. The potential significance of space sciences to the human cognitive environment is yet to be fully ascertained. We now understand that the entire history of the universe informs each and every particle and spin of the fabric of existence. The implications of this knowledge have the power to facilitate our overcoming many social diseases such as racism, nationalism and the ideological delusions that tolerate such activities as warfare. Space sciences may help to purge the human cognitive atmosphere of those ills and ignorance that sap global resources, challenging global sustainability, from the economic to the psychosocial. Were the full implications of our united origins and destiny as a cosmic organism to be applied to how we live as a species on the Earth, there would be adequate funds for all manner of science and education such as to transform the global human and ecological landscape in ways as yet only dreamt or fictionalized. The bioethics of universal
-
International Nuclear Information System (INIS)
Hirano, Kunio; Konagaya, Shuhei; Turner, Alexander; Noda, Yuichiro; Kitamura, Shigeru; Kotera, Hidetoshi; Iwata, Hiroo
2017-01-01
Human pluripotent stem cells (hPSCs) are thought to be a promising cell-source solution for regenerative medicine due to their indefinite proliferative potential and ability to differentiate to functional somatic cells. However, issues remain with regard to achieving reproducible differentiation of cells with the required functionality for realizing human transplantation therapies and with regard to reducing the potential for bacterial or fungal contamination. To meet these needs, we have developed a closed-channel culture device and corresponding control system. Uniformly-sized spheroidal hPSCs aggregates were formed inside wells within a closed-channel and maintained continuously throughout the culture process. Functional islet-like endocrine cell aggregates were reproducibly induced following a 30-day differentiation protocol. Our system shows an easily scalable, novel method for inducing PSC differentiation with both purity and functionality. - Highlights: • A simple, closed-channel-based, semi-automatic culture system is proposed. • Uniform cell aggregate formation and culture is realized in microwell structure. • Functional islet cells are successfully induced following 30-plus-day protocol. • System requires no daily medium replacement and reduces contamination risk.
-
International Nuclear Information System (INIS)
Dracott, B.N.
1974-01-01
Cell viability and the production of direct PFC were studied in mouse spleen cell cultures after cortisol treatment in vivo or in vitro at various times relative to primary stimulation with SRBC in vitro. Cortisol treatment in vivo reduced spleen cell numbers by 88 percent after 48 hr, but cultures of the remaining cells produced as many PFC in vitro as did cultures of equal numbers of normal spleen cells. In normal spleen cell cultures incubated with cortisol for 4 hr prior to the addition of antigen, peak responses of PFC/culture and PFC/10 6 cells occurred 24 hr later than in controls and averaged, respectively, 27 and 141 percent of control values. Minimum viable cell numbers were observed in cortisol-treated cultures after 3 days; thereafter cell numbers gradually increased. These results were not significantly altered when cultures were treated simultaneously with cortisol and antigen. The response was not suppressed if the addition of antigen preceded that of cortisol by more than 4 hr. Suppression was also considerably reduced if fetal calf serum was used when preparing cells for culture
-
Rennert, Knut; Nitschke, Mirko; Wallert, Maria; Keune, Natalie; Raasch, Martin; Lorkowski, Stefan; Mosig, Alexander S
2017-01-01
Harvesting cultivated macrophages for tissue engineering purposes by enzymatic digestion of cell adhesion molecules can potentially result in unintended activation, altered function, or behavior of these cells. Thermo-responsive polymer is a promising tool that allows for gentle macrophage detachment without artificial activation prior to subculture within engineered tissue constructs. We therefore characterized different species of thermo-responsive polymers for their suitability as cell substrate and to mediate gentle macrophage detachment by temperature shift. Primary human monocyte- and THP-1-derived macrophages were cultured on thermo-responsive polymers and characterized for phagocytosis and cytokine secretion in response to lipopolysaccharide stimulation. We found that both cell types differentially respond in dependence of culture and stimulation on thermo-responsive polymers. In contrast to THP-1 macrophages, primary monocyte-derived macrophages showed no signs of impaired viability, artificial activation, or altered functionality due to culture on thermo-responsive polymers compared to conventional cell culture. Our study demonstrates that along with commercially available UpCell carriers, two other thermo-responsive polymers based on poly(vinyl methyl ether) blends are attractive candidates for differentiation and gentle detachment of primary monocyte-derived macrophages. In summary, we observed similar functionality and viability of primary monocyte-derived macrophages cultured on thermo-responsive polymers compared to standard cell culture surfaces. While this first generation of custom-made thermo-responsive polymers does not yet outperform standard culture approaches, our results are very promising and provide the basis for exploiting the unique advantages offered by custom-made thermo-responsive polymers to further improve macrophage culture and recovery in the future, including the covalent binding of signaling molecules and the reduction of
-
Influence of physicochemical properties of beryllium particles on cultured cell toxicity
International Nuclear Information System (INIS)
Finch, G.L.; Brooks, A.L.; Hoover, M.D.; Cuddihy, R.G.
1988-01-01
The toxicity of beryllium oxide (BeO)), beryllium metal, and beryllium sulfate (BeSO 4 ) was studied in two cell lines, Chinese hamster ovary cells (CHO) and lung epithelial cells (LEC). Beryllium oxide particles were prepared at either 500 or 1000 deg. C, and two different particle sizes of beryllium metal were used. Following a 20-h exposure to beryllium compounds, cells were grown in culture to quantitate cloning ability relative to controls as a measure of cell killing, The LEC cultures were more sensitive to beryllium cytotoxicity than the CHO cells. When expressed on the basis of the mass of material added to the cultures, the order of toxicity was BeSO 4 ≥ 500 deg. C -BeO > 1000 deg. C -BeO > Be metal (small) Be metal (large). When cytotoxic effects were expressed on the basis of particulate surface rather than mass, the relative differences in toxicity between compounds was decreased. The order of toxicity was Be metal (small) ∼ Be metal (large) ∼ 500 deg. C-BeO ∼ 1000 deg. C-BeO. These data indicate that solubility influences beryllium toxicity to short-term cell cultures. (author)
-
Establishment of three-dimensional cultures of human pancreatic duct epithelial cells
International Nuclear Information System (INIS)
Gutierrez-Barrera, Angelica M.; Menter, David G.; Abbruzzese, James L.; Reddy, Shrikanth A.G.
2007-01-01
Three-dimensional (3D) cultures of epithelial cells offer singular advantages for studies of morphogenesis or the role of cancer genes in oncogenesis. In this study, as part of establishing a 3D culture system of pancreatic duct epithelial cells, we compared human pancreatic duct epithelial cells (HPDE-E6E7) with pancreatic cancer cell lines. Our results show, that in contrast to cancer cells, HPDE-E6E7 organized into spheroids with what appeared to be apical and basal membranes and a luminal space. Immunostaining experiments indicated that protein kinase Akt was phosphorylated (Ser473) and CTMP, a negative Akt regulator, was expressed in both HPDE-E6E7 and cancer cells. However, a nuclear pool of CTMP was detectable in HPDE-E6E7 cells that showed a dynamic concentrated expression pattern, a feature that further distinguished HPDE-E637 cells from cancer cells. Collectively, these data suggest that 3D cultures of HPDE-E6E7 cells are useful for investigating signaling and morphological abnormalities in pancreatic cancer cells
-
Properties of Dental Pulp-derived Mesenchymal Stem Cells and the Effects of Culture Conditions.
Kawashima, Nobuyuki; Noda, Sonoko; Yamamoto, Mioko; Okiji, Takashi
2017-09-01
Dental pulp mesenchymal stem cells (DPMSCs) highly express mesenchymal stem cell markers and possess the potential to differentiate into neural cells, osteoblasts, adipocytes, and chondrocytes. Thus, DPMSCs are considered suitable for tissue regeneration. The colony isolation method has commonly been used to collect relatively large amounts of heterogeneous DPMSCs. Homogenous DPMSCs can be isolated by fluorescence-activated cell sorting using antibodies against mesenchymal stem cell markers, although this method yields a limited number of cells. Both quality and quantity of DPMSCs are critical to regenerative therapy, and cell culture methods need to be improved. We thus investigated the properties of DPMSCs cultured with different methods. DPMSCs in a three-dimensional spheroid culture system, which is similar to the hanging drop culture for differentiation of embryonic stem cells, showed upregulation of odonto-/osteoblastic markers and mineralized nodule formation. This suggests that this three-dimensional spheroid culturing system for DPMSCs may be suitable for inducing hard tissues. We further examined the effect of cell culture density on the properties of DPMSCs because the properties of stem cells can be altered depending on the cell density. DPMSCs cultured under the confluent cell density condition showed slight downregulation of some mesenchymal stem cell markers compared with those under the sparse condition. The ability of DPMSCs to differentiate into hard tissue-forming cells was found to be enhanced in the confluent condition, suggesting that the confluent culture condition may not be suitable for maintaining the stemness of DPMSCs. When DPMSCs are to be used for hard tissue regeneration, dense followed by sparse cell culture conditions may be a better alternative strategy. Copyright © 2017 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.
-
Radiation transformation in differentiated human cells in culture
International Nuclear Information System (INIS)
Mothersill, C.; Seymour, C.; Moriarty, M.; Malone, J.; Byrne, P.; Hennessy, T.
1986-01-01
A tissue culture technique is described for human thyroid tissue as an approach to studying mechanisms of human radiation carcinogenesis. Normal human tissue obtained from surgery is treated in one of two ways, depending upon size of specimen. Large pieces are completely digested in trypsin/ collagenase solution to a single cell suspension. Small pieces of tissue are plated as explants following partial digestion in trypsin/collagenase solution. Following irradiation of the primary differentiated monolayers (normally 10 days after plating), the development of transformed characteristics is monitored in the subsequent subcultures. A very high level of morphological and functional differentiation is apparent in the primary cultures. Over a period of approx. 6 months, the irradiated surviving cells continue to grow in culture, unlike the unirradiated controls which senesce after 2-3 subcultures. (UK)
-
Immunophenotype of hematopoietic stem cells from placental/umbilical cord blood after culture
Directory of Open Access Journals (Sweden)
P. Pranke
2005-12-01
Full Text Available Identification and enumeration of human hematopoietic stem cells remain problematic, since in vitro and in vivo stem cell assays have different outcomes. We determined if the altered expression of adhesion molecules during stem cell expansion could be a reason for the discrepancy. CD34+CD38- and CD34+CD38+ cells from umbilical cord blood were analyzed before and after culture with thrombopoietin (TPO, FLT-3 ligand (FL and kit ligand (KL; or stem cell factor in different combinations: TPO + FL + KL, TPO + FL and TPO, at concentrations of 50 ng/mL each. Cells were immunophenotyped by four-color fluorescence using antibodies against CD11c, CD31, CD49e, CD61, CD62L, CD117, and HLA-DR. Low-density cord blood contained 1.4 ± 0.9% CD34+ cells, 2.6 ± 2.1% of which were CD38-negative. CD34+ cells were isolated using immuno-magnetic beads and cultured for up to 7 days. The TPO + FL + KL combination presented the best condition for maintenance of stem cells. The total cell number increased 4.3 ± 1.8-fold, but the number of viable CD34+ cells decreased by 46 ± 25%. On the other hand, the fraction of CD34+CD38- cells became 52.0 ± 29% of all CD34+ cells. The absolute number of CD34+CD38- cells was expanded on average 15 ± 12-fold when CD34+ cells were cultured with TPO + FL + KL for 7 days. The expression of CD62L, HLA-DR and CD117 was modulated after culture, particularly with TPO + FL + KL, explaining differences between the adhesion and engraftment of primary and cultured candidate stem cells. We conclude that culture of CD34+ cells with TPO + FL + KL results in a significant increase in the number of candidate stem cells with the CD34+CD38- phenotype.
-
Yen, Meng-Hua; Wu, Yuan-Yi; Liu, Yi-Shiuan; Rimando, Marilyn; Ho, Jennifer Hui-Chun; Lee, Oscar Kuang-Sheng
2016-08-19
Mesenchymal stromal cells (MSCs) are multipotent and have great potential in cell therapy. Previously we reported the differentiation potential of human MSCs into hepatocytes in vitro and that these cells can rescue fulminant hepatic failure. However, the conventional static culture method neither maintains growth factors at an optimal level constantly nor removes cellular waste efficiently. In addition, not only is the duration of differentiating hepatocyte lineage cells from MSCs required to improve, but also the need for a large number of hepatocytes for cell therapy has not to date been addressed fully. The purpose of this study is to design and develop an innovative microfluidic device to overcome these shortcomings. We designed and fabricated a microfluidic device and a culture system for hepatic differentiation of MSCs using our protocol reported previously. The microfluidic device contains a large culture chamber with a stable uniform flow to allow homogeneous distribution and expansion as well as efficient induction of hepatic differentiation for MSCs. The device enables real-time observation under light microscopy and exhibits a better differentiation efficiency for MSCs compared with conventional static culture. MSCs grown in the microfluidic device showed a higher level of hepatocyte marker gene expression under hepatic induction. Functional analysis of hepatic differentiation demonstrated significantly higher urea production in the microfluidic device after 21 days of hepatic differentiation. The microfluidic device allows the generation of a large number of MSCs and induces hepatic differentiation of MSCs efficiently. The device can be adapted for scale-up production of hepatic cells from MSCs for cellular therapy.
-
Six cloned calves produced from adult fibroblast cells after long-term culture
Kubota, Chikara; Yamakuchi, Hiroshi; Todoroki, Junichi; Mizoshita, Kazunori; Tabara, Norio; Barber, Michele; Yang, Xiangzhong
2000-01-01
Cloning whole animals with somatic cells as parents offers the possibility of targeted genetic manipulations in vitro such as “gene knock-out” by homologous recombination. However, such manipulation requires prolonged culture of nuclear donor cells. Previous successes in cloning have been limited to the use of cells collected either fresh or after short-term culture. Therefore, demonstration of genetic totipotency of cells after prolonged culture is pivotal to combining site-specific genetic manipulations and cloning. Here we report birth of six clones of an aged (17-year-old) Japanese Black Beef bull using ear skin fibroblast cells as nuclear donor cells after up to 3 months of in vitro culture (10–15 passages). We observed higher developmental rates for embryos derived from later passages (10 and 15) as compared with those embryos from an early passage (passage 5). The four surviving clones are now 10–12 months of age and appear normal, similar to their naturally reproduced peers. These data show that fibroblasts of aged animals remain competent for cloning, and prolonged culture does not affect the cloning competence of adult somatic donor cells. PMID:10655472
-
Sequential cancer mutations in cultured human intestinal stem cells
Drost, Jarno; van Jaarsveld, Richard H.; Ponsioen, Bas; Zimberlin, Cheryl; van Boxtel, Ruben; Buijs, Arjan; Sachs, Norman; Overmeer, René M.; Offerhaus, G. Johan; Begthel, Harry; Korving, Jeroen; van de Wetering, Marc; Schwank, Gerald; Logtenberg, Meike; Cuppen, Edwin; Snippert, Hugo J.; Medema, Jan Paul; Kops, Geert J. P. L.; Clevers, Hans
2015-01-01
Crypt stem cells represent the cells of origin for intestinal neoplasia. Both mouse and human intestinal stem cells can be cultured in medium containing the stem-cell-niche factors WNT, R-spondin, epidermal growth factor (EGF) and noggin over long time periods as epithelial organoids that remain
-
Directory of Open Access Journals (Sweden)
Dai Fei Elmer Ker
Full Text Available Current cell culture practices are dependent upon human operators and remain laborious and highly subjective, resulting in large variations and inconsistent outcomes, especially when using visual assessments of cell confluency to determine the appropriate time to subculture cells. Although efforts to automate cell culture with robotic systems are underway, the majority of such systems still require human intervention to determine when to subculture. Thus, it is necessary to accurately and objectively determine the appropriate time for cell passaging. Optimal stem cell culturing that maintains cell pluripotency while maximizing cell yields will be especially important for efficient, cost-effective stem cell-based therapies. Toward this goal we developed a real-time computer vision-based system that monitors the degree of cell confluency with a precision of 0.791±0.031 and recall of 0.559±0.043. The system consists of an automated phase-contrast time-lapse microscope and a server. Multiple dishes are sequentially imaged and the data is uploaded to the server that performs computer vision processing, predicts when cells will exceed a pre-defined threshold for optimal cell confluency, and provides a Web-based interface for remote cell culture monitoring. Human operators are also notified via text messaging and e-mail 4 hours prior to reaching this threshold and immediately upon reaching this threshold. This system was successfully used to direct the expansion of a paradigm stem cell population, C2C12 cells. Computer-directed and human-directed control subcultures required 3 serial cultures to achieve the theoretical target cell yield of 50 million C2C12 cells and showed no difference for myogenic and osteogenic differentiation. This automated vision-based system has potential as a tool toward adaptive real-time control of subculturing, cell culture optimization and quality assurance/quality control, and it could be integrated with current and
-
Lipoprotein receptors in cultured bovine endothelial cells
International Nuclear Information System (INIS)
Struempfer, A.E.M.
1983-07-01
In this study, receptors that may be involved in the uptake of low density lipoproteins (LDL) and low density lipoproteins which have been modified by acetylation (AcLDL), were characterized. Aortic epithelial cells were used and a cell culture system which closely resembled the in vivo monolayer was established. Endothelial cell and lipoprotein interactions were examined by incubating the cells with 125 l-labelled lipoproteins under various conditions. The receptor affinity of bovine aortic endothelial cells was higher for AcLDL than that for LDL. Competition studies demonstrated that there were two distinct receptors for LDL and AcLDL on the endothelial cells. AcLDL did not compete with LDL for the LDL receptor, and conversely LDL did not compete with AcLDL for the AcLDL receptor. The receptor activities for LDL and AcLDL were examined as a function of culture age. Whereas the LDL receptor could be regulated, the AcLDL receptor was not as susceptible to regulation. Upon exposing endothelial cells for 72 h to either LDL or AcLDL, it was found that the total amount of cellular cholesterol increased by about 50%. However, the increase of total cholesterol was largely in the form of free cholesterol. This is in contrast to macrophages, where the increase in total cholesterol upon exposure to AcLDL is largely in the form cholesteryl esters
-
Ethical and cultural value of the Earth sciences. Interview with Prof. Giulio Giorello
Directory of Open Access Journals (Sweden)
Silvia Peppoloni
2012-07-01
Full Text Available Prof. Giulio Giorello is amongst the most prominent philosophers of science in Italy and in the world. He is currently Professor of the Philosophy of Science at the University of Milan, Director of the Series ‘Science and Ideas’ (Raffaele Cortina Books Editor, and Literary Journalist of the cultural pages of the Corriere della Sera, one of the most important of the Italian newspapers. In this keynote presentation, in interview form, he talks about the value that the Earth sciences have had through history, framing this group of disciplines in ethical and epistemological terms, and highlighting some important elements that have to be considered in geological activities.
-
Race and Ethnicity: Powerful Cultural Forecasters of Science Learning and Performance
Atwater, Mary M.; Lance, Jennifer; Woodard, UrLeaka; Johnson, Natasha Hillsman
2013-01-01
This article addresses the impact of race and ethnicity on students' science learning in US schools. Specifically, it discusses (a) the constructs of race, ethnicity, and culture, and the racial and ethnic student composition in US public schools; (b) effective classroom practices for curriculum, instruction, and assessment related to race…
-
Allon, Aliza A; Schneider, Richard A; Lotz, Jeffrey C
2009-01-01
Our goal is to optimize stem cell-based tissue engineering strategies in the context of the intervertebral disc environment. We explored the benefits of co-culturing nucleus pulposus cells (NPC) and adult mesenchymal stem cells (MSC) using a novel spherical bilaminar pellet culture system where one cell type is enclosed in a sphere of the other cell type. Our 3D system provides a structure that exploits embryonic processes such as tissue induction and condensation. We observed a unique phenomenon: the budding of co-culture pellets and the formation of satellite pellets that separate from the main pellet. MSC and NPC co-culture pellets were formed with three different structural organizations. The first had random organization. The other two had bilaminar organization with either MSC inside and NPC outside or NPC inside and MSC outside. By 14 days, all co-culture pellets exhibited budding and spontaneously generated satellite pellets. The satellite pellets were composed of both cell types and, surprisingly, all had the same bilaminar organization with MSC on the inside and NPC on the outside. This organization was independent of the structure of the main pellet that the satellites stemmed from. The main pellets generated satellite pellets that spontaneously organized into a bilaminar structure. This implies that structural organization occurs naturally in this cell culture system and may be inherently favorable for cell-based tissue engineering strategies. The occurrence of budding and the organization of satellite pellets may have important implications for the use of co-culture pellets in cell-based therapies for disc regeneration. From a therapeutic point of view, the generation of satellite pellets may be a beneficial feature that would serve to spread donor cells throughout the host matrix and restore normal matrix composition in a sustainable way, ultimately renewing tissue function.
-
Animal-cell culture in aqueous two-phase systems
Zijlstra, G.M.
1998-01-01
In current industrial biotechnology, animal-cell culture is an important source of therapeutic protein products. The conventional animal-cell production processes, however, include many unit operations as part of the fermentation and downstream processing strategy. The research described in