WorldWideScience

Sample records for 3d culture implications

  1. 3D culture for cardiac cells.

    Science.gov (United States)

    Zuppinger, Christian

    2016-07-01

    This review discusses historical milestones, recent developments and challenges in the area of 3D culture models with cardiovascular cell types. Expectations in this area have been raised in recent years, but more relevant in vitro research, more accurate drug testing results, reliable disease models and insights leading to bioartificial organs are expected from the transition to 3D cell culture. However, the construction of organ-like cardiac 3D models currently remains a difficult challenge. The heart consists of highly differentiated cells in an intricate arrangement.Furthermore, electrical “wiring”, a vascular system and multiple cell types act in concert to respond to the rapidly changing demands of the body. Although cardiovascular 3D culture models have been predominantly developed for regenerative medicine in the past, their use in drug screening and for disease models has become more popular recently. Many sophisticated 3D culture models are currently being developed in this dynamic area of life science. This article is part of a Special Issue entitled: Cardiomyocyte Biology: Integration of Developmental and Environmental Cues in the Heart edited by Marcus Schaub and Hughes Abriel.

  2. 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.

  3. Long-Term Cultures of Human Cornea Limbal Explants Form 3D Structures Ex Vivo - Implications for Tissue Engineering and Clinical Applications.

    Science.gov (United States)

    Szabó, Dóra Júlia; Noer, Agate; Nagymihály, Richárd; Josifovska, Natasha; Andjelic, Sofija; Veréb, Zoltán; Facskó, Andrea; Moe, Morten C; Petrovski, Goran

    2015-01-01

    Long-term cultures of cornea limbal epithelial stem cells (LESCs) were developed and characterized for future tissue engineering and clinical applications. The limbal tissue explants were cultivated and expanded for more than 3 months in medium containing serum as the only growth supplement and without use of scaffolds. Viable 3D cell outgrowth from the explants was observed within 4 weeks of cultivation. The outgrowing cells were examined by immunofluorescent staining for putative markers of stemness (ABCG2, CK15, CK19 and Vimentin), proliferation (p63α, Ki-67), limbal basal epithelial cells (CK8/18) and differentiated cornea epithelial cells (CK3 and CK12). Morphological and immunostaining analyses revealed that long-term culturing can form stratified 3D tissue layers with a clear extracellular matrix deposition and organization (collagen I, IV and V). The LESCs showed robust expression of p63α, ABCG2, and their surface marker fingerprint (CD117/c-kit, CXCR4, CD146/MCAM, CD166/ALCAM) changed over time compared to short-term LESC cultures. Overall, we provide a model for generating stem cell-rich, long-standing 3D cultures from LESCs which can be used for further research purposes and clinical transplantation.

  4. Filling gaps in cultural heritage documentation by 3D photography

    Directory of Open Access Journals (Sweden)

    W. Schuhr

    2015-08-01

    Niepce (1827, but seem to promise a great future also in 3D Cultural Heritage documentation. *Last not least 3D printers more and more seem to conquer the IT-market, obviously showing an international competition.

  5. Multizone Paper Platform for 3D Cell Cultures

    Science.gov (United States)

    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

  6. Beyond 3D culture models of cancer

    Science.gov (United States)

    Tanner, Kandice; Gottesman, Michael M.

    2016-01-01

    The mechanisms underlying the spatiotemporal evolution of tumor ecosystems present a challenge in evaluating drug efficacy. In this Perspective, we address the use of three-dimensional in vitro culture models to delineate the dynamic interplay between the tumor and the host microenvironment in an effort to attain realistic platforms for assessing pharmaceutical efficacy in patients. PMID:25877888

  7. Filling gaps in cultural heritage documentation by 3D photography

    Science.gov (United States)

    Schuhr, W.; Lee, J. D.

    2015-08-01

    This contribution promotes 3D photography as an important tool to obtain objective object information. Keeping mainly in mind World Heritage documentation as well as Heritage protection, it is another intention of this paper, to stimulate the interest in applications of 3D photography for professionals as well as for amateurs. In addition this is also an activity report of the international CIPA task group 3. The main part of this paper starts with "Digging the treasure of existing international 3D photography". This does not only belong to tangible but also to intangible Cultural Heritage. 3D photography clearly supports the recording, the visualization, the preservation and the restoration of architectural and archaeological objects. Therefore the use of 3D photography in C.H. should increase on an international level. The presented samples in 3D represent a voluminous, almost partly "forgotten treasure" of international archives for 3D photography. The next chapter is on "Promoting new 3D photography in Cultural Heritage". Though 3D photographs are a well-established basic photographic and photogrammetric tool, even suited to provide "near real" documentation, they are still a matter of research and improvement. Beside the use of 3D cameras even single lenses cameras are very much suited for photographic 3D documentation purposes in Cultural Heritage. Currently at the Faculty of Civil Engineering of the University of Applied Sciences Magdeburg-Stendal, low altitude aerial photography is exposed from a maximum height of 13m, using a hand hold carbon telescope rod. The use of this "huge selfie stick" is also an (international) recommendation, to expose high resolution 3D photography of monuments under expedition conditions. In addition to the carbon rod recently a captive balloon and a hexacopter UAV- platform is in use, mainly to take better synoptically (extremely low altitude, ground truth) aerial photography. Additional experiments with respect to "easy

  8. Dynamic 3D cell culture via a chemoselective photoactuated ligand.

    Science.gov (United States)

    Westcott, Nathan P; Luo, Wei; Goldstein, Jeffrey; Yousaf, Muhammad N

    2014-09-01

    A new strategy to create a dynamic scaffold for three-dimensional (3D) cell experiments based on a photo-activated cell adhesive peptide ligand is described. After polymerization, the inert matrix becomes cell adhesive by chemoselective modification through the conjugation of oxyamine-terminated ligands. Furthermore, spatial and temporal control of cell culture within the 3D matrix was achieved by the use of a biospecific photoprotected peptide and visualized by confocal microscopy.

  9. Fabrication of Nanostructured Poly-ε-caprolactone 3D Scaffolds for 3D Cell Culture Technology

    KAUST Repository

    Schipani, Rossana

    2015-04-21

    Tissue engineering is receiving tremendous attention due to the necessity to overcome the limitations related to injured or diseased tissues or organs. It is the perfect combination of cells and biomimetic-engineered materials. With the appropriate biochemical factors, it is possible to develop new effective bio-devices that are capable to improve or replace biological functions. Latest developments in microfabrication methods, employing mostly synthetic biomaterials, allow the production of three-dimensional (3D) scaffolds that are able to direct cell-to-cell interactions and specific cellular functions in order to drive tissue regeneration or cell transplantation. The presented work offers a rapid and efficient method of 3D scaffolds fabrication by using optical lithography and micro-molding techniques. Bioresorbable polymer poly-ε-caprolactone (PCL) was the material used thanks to its high biocompatibility and ability to naturally degrade in tissues. 3D PCL substrates show a particular combination in the designed length scale: cylindrical shaped pillars with 10μm diameter, 10μm height, arranged in a hexagonal lattice with spacing of 20μm were obtained. The sidewalls of the pillars were nanostructured by attributing a 3D architecture to the scaffold. The suitability of these devices as cell culture technology supports was evaluated by plating NIH/3T3 mouse embryonic fibroblasts and human Neural Stem Cells (hNSC) on them. Scanning Electron Microscopy (SEM) analysis was carried out in order to examine the micro- and nano-patterns on the surface of the supports. In addition, after seeding of cells, SEM and immunofluorescence characterization of the fabricated systems were performed to check adhesion, growth and proliferation. It was observed that cells grow and develop healthy on the bio-polymeric devices by giving rise to well-interconnected networks. 3D PCL nano-patterned pillared scaffold therefore may have considerable potential as effective tool for

  10. Nanomagnetic Levitation 3-D Cultures of Breast and Colorectal Cancers

    Science.gov (United States)

    Bumpers, Harvey L.; Janagama, Dasharatham G.; Manne, Upender; Basson, Marc D.; Katkoori, Venkat

    2014-01-01

    Background Innovative technologies for drug discovery and development, cancer models, stem cell research, tissue engineering, and drug testing in various cell-based platforms require an application similar to the in vivo system. Materials and Methods We developed for the first time nanomagnetically levitated three dimensional (3-D) cultures of breast cancer (BC) and colorectal cancer (CRC) cells using carbon encapsulated cobalt magnetic nanoparticles. BC and CRC xenografts grown in severe combined immunodeficient (SCID) mice were evaluated for N-cadherin and Epidermal growth factor receptor (EGFR) expressions. These phenotypes were compared with 2-D cultures and 3-D cultures grown in a gel matrix. Results The BC and CRC cells grown by magnetic levitation formed microtissues. The levitated cultures had high viability and were maintained in culture for long periods of time. It has been observed that N-cadherin and EGFR activities were highly expressed in the levitated 3-D tumor spheres and xenografts of CRC and BC cells. Conclusions Nanomagnetically levitated 3-D cultures tend to form stable microtissues of BC and CRC and may be more feasible for a range of applications in drug discovery or regenerative medicine. PMID:25617973

  11. An Optically Controlled 3D Cell Culturing System

    Directory of Open Access Journals (Sweden)

    Kelly S. Ishii

    2011-01-01

    Full Text Available A novel 3D cell culture system was developed and tested. The cell culture device consists of a microfluidic chamber on an optically absorbing substrate. Cells are suspended in a thermoresponsive hydrogel solution, and optical patterns are utilized to heat the solution, producing localized hydrogel formation around cells of interest. The hydrogel traps only the desired cells in place while also serving as a biocompatible scaffold for supporting the cultivation of cells in 3D. This is demonstrated with the trapping of MDCK II and HeLa cells. The light intensity from the optically induced hydrogel formation does not significantly affect cell viability.

  12. 2D- and 3D-culture of cell

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    Khoruzhenko A. I.

    2011-02-01

    Full Text Available The cultivation of mammalian cells in three-dimensional conditions acquires a priority in a variety of biomedical applications. In the areas of toxicology and anticancer drug development it concerns a significant difference of responses to proapoptotic factors of the cells cultured in 2D versus 3D environment. Besides, the clear-cut differences have been found in cell polarity, cytoskeleton structure, distribution of receptors to wide range of hormones, growth factors, etc. in mammalian cells depending on culture conditions. It is resulted in different response of cultured cells to extracellular stimuli. Multicellular spheroids are regarded presently as the most convenient model of solid tumour growth in vitro. The cultivation of thyroid follicles, mammary acini and other structure units, maintaining initial tissue organization, allows studying the behavior, biochemical features and gene profile of differentiated cells. On the other hand, 3D cultures have some limitations in comparison with a well established monolayer culture. The advantages and disadvantages of each type of cultures and their application in biological and medical researches will be discussed in this review

  13. Single molecule microscopy in 3D cell cultures and tissues.

    Science.gov (United States)

    Lauer, Florian M; Kaemmerer, Elke; Meckel, Tobias

    2014-12-15

    From the onset of the first microscopic visualization of single fluorescent molecules in living cells at the beginning of this century, to the present, almost routine application of single molecule microscopy, the method has well-proven its ability to contribute unmatched detailed insight into the heterogeneous and dynamic molecular world life is composed of. Except for investigations on bacteria and yeast, almost the entire story of success is based on studies on adherent mammalian 2D cell cultures. However, despite this continuous progress, the technique was not able to keep pace with the move of the cell biology community to adapt 3D cell culture models for basic research, regenerative medicine, or drug development and screening. In this review, we will summarize the progress, which only recently allowed for the application of single molecule microscopy to 3D cell systems and give an overview of the technical advances that led to it. While initially posing a challenge, we finally conclude that relevant 3D cell models will become an integral part of the on-going success of single molecule microscopy.

  14. Molecular predictors of 3D morphogenesis by breast cancer cell lines in 3D culture.

    Directory of Open Access Journals (Sweden)

    Ju Han

    2010-02-01

    Full Text Available Correlative analysis of molecular markers with phenotypic signatures is the simplest model for hypothesis generation. In this paper, a panel of 24 breast cell lines was grown in 3D culture, their morphology was imaged through phase contrast microscopy, and computational methods were developed to segment and represent each colony at multiple dimensions. Subsequently, subpopulations from these morphological responses were identified through consensus clustering to reveal three clusters of round, grape-like, and stellate phenotypes. In some cases, cell lines with particular pathobiological phenotypes clustered together (e.g., ERBB2 amplified cell lines sharing the same morphometric properties as the grape-like phenotype. Next, associations with molecular features were realized through (i differential analysis within each morphological cluster, and (ii regression analysis across the entire panel of cell lines. In both cases, the dominant genes that are predictive of the morphological signatures were identified. Specifically, PPARgamma has been associated with the invasive stellate morphological phenotype, which corresponds to triple-negative pathobiology. PPARgamma has been validated through two supporting biological assays.

  15. Molecular Predictors of 3D Morphogenesis by Breast Cancer Cell Lines in 3D Culture

    Energy Technology Data Exchange (ETDEWEB)

    Han, Ju; Chang, Hang; Giricz, Orsi; Lee, Genee; Baehner, Frederick; Gray, Joe; Bissell, Mina; Kenny, Paraic; Parvin, Bahram

    2010-02-01

    Correlative analysis of molecular markers with phenotypic signatures is the simplest model for hypothesis generation. In this paper, a panel of 24 breast cell lines was grown in 3D culture, their morphology was imaged through phase contrast microscopy, and computational methods were developed to segment and represent each colony at multiple dimensions. Subsequently, subpopulations from these morphological responses were identified through consensus clustering to reveal three clusters of round, grape-like, and stellate phenotypes. In some cases, cell lines with particular pathobiological phenotypes clustered together (e.g., ERBB2 amplified cell lines sharing the same morphometric properties as the grape-like phenotype). Next, associations with molecular features were realized through (i) differential analysis within each morphological cluster, and (ii) regression analysis across the entire panel of cell lines. In both cases, the dominant genes that are predictive of the morphological signatures were identified. Specifically, PPAR? has been associated with the invasive stellate morphological phenotype, which corresponds to triple-negative pathobiology. PPAR? has been validated through two supporting biological assays.

  16. Ideal Positions: 3D Sonography, Medical Visuality, Popular Culture.

    Science.gov (United States)

    Seiber, Tim

    2016-03-01

    As digital technologies are integrated into medical environments, they continue to transform the experience of contemporary health care. Importantly, medicine is increasingly visual. In the history of sonography, visibility has played an important role in accessing fetal bodies for diagnostic and entertainment purposes. With the advent of three-dimensional (3D) rendering, sonography presents the fetus visually as already a child. The aesthetics of this process and the resulting imagery, made possible in digital networks, discloses important changes in the relationship between technology and biology, reproductive health and political debates, and biotechnology and culture.

  17. Surface modified alginate microcapsules for 3D cell culture

    Science.gov (United States)

    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.

  18. Platelet gel: 3D scaffold for cell culture

    OpenAIRE

    Andrei Moroz; Renata Aparecida de Camargo Bittencourt; Sérgio Luis Felisbino; Hamilton da Rosa Pereira; Rosana Rossi-Ferreira; Elenice Deffune

    2009-01-01

    INTRODUÇÃO: O reparo tissular é o objetivo final da cirurgia. A cultura celular requer arcabouço mecânico que dê suporte ao crescimento celular e difusão dos nutrientes. O uso do plasma rico em plaquetas (PRP) como um arcabouço 3D possui diversas vantagens: é material biológico, de fácil absorção pós-transplante, rico em fatores de crescimento, em especial PDGF- ββ e TGF-β que estimula síntese de matriz extracelular na cartilagem. OBJETIVO: Desenvolver arcabouço 3D à base de PR...

  19. Gel de plaquetas: arcabouço 3D para cultura celular Platelet gel: 3D scaffold for cell culture

    Directory of Open Access Journals (Sweden)

    Andrei Moroz

    2009-01-01

    Full Text Available INTRODUÇÃO: O reparo tissular é o objetivo final da cirurgia. A cultura celular requer arcabouço mecânico que dê suporte ao crescimento celular e difusão dos nutrientes. O uso do plasma rico em plaquetas (PRP como um arcabouço 3D possui diversas vantagens: é material biológico, de fácil absorção pós-transplante, rico em fatores de crescimento, em especial PDGF- ββ e TGF-β que estimula síntese de matriz extracelular na cartilagem. OBJETIVO: Desenvolver arcabouço 3D à base de PRP. MATERIAIS E MÉTODOS: Duas formas foram idealizadas: Sphere e Carpet. Condições estéreis foram utilizadas. O gel de plaquetas permaneceu em cultura celular, observado diariamente em microscópio invertido. RESULTADOS: Ambos arcabouços obtiveram sucesso, com aspectos positivos e negativos. DISCUSSÃO: A forma Sphere não aderiu ao plástico. Observou-se retração do gel e investigação ao microscópio dificultada devido às áreas opacas no campo visual. A forma Carpet não aderiu ao plástico e apresentou-se translúcida. O tempo de estudo foi de 20 dias. CONCLUSÕES: A produção de um arcabouço 3D PRP foi um sucesso, e trata-se de uma alternativa que necessita ser mais utilizado e investigado para que se consolide em uma rota eficiente e confiável na tecnologia de engenharia tissular, particularmente em cultura de tecido cartilaginoso.INTRODUCTION: Tissue repair has been the ultimate goal of surgery. Cell culture requires a mechanical scaffold that supports cell growth and nutrient diffusion. Using platelet-rich plasma (PRP as a 3D scaffold presents various advantages: it is a biological material, easily absorbed after transplantation, rich in growth factors, in particular, PDGF-ββ and TGF-β that stimulate extracellular matrix synthesis in cartilage culture. OBJECTIVE: To develop a PRP 3D scaffold. Material and METHODS: Two forms were idealized: Sphere and Carpet. Sterile conditions were used. The platelet gel remained in culture

  20. Impedance Spectroscopic Characterisation of Porosity in 3D Cell Culture Scaffolds with Different Channel Networks

    DEFF Research Database (Denmark)

    Canali, Chiara; Mohanty, Soumyaranjan; Heiskanen, Arto

    2015-01-01

    We present the application of electrochemical impedance spectroscopy (EIS) as a method for discriminating between different polydimethylsiloxane (PDMS) scaffolds for three-dimensional (3D) cell cultures. The validity of EIS characterisation for scaffolds having different degree of porosity...... serve as means of single-frequency measurements for fast scaffold characterization combined with in vitro monitoring of 3D cell cultures....

  1. Design of 3D printed insert for hanging culture of Caco-2 cells.

    Science.gov (United States)

    Shen, Chong; Meng, Qin; Zhang, Guoliang

    2014-12-17

    A Caco-2 cell culture on Transwell, an alternative testing to animal or human testing used in evaluating drug intestinal permeability, incorrectly estimated the absorption of actively transported drugs due to the low expression of membrane transporters. Similarly, three-dimensional (3D) cultures of Caco-2 cells, which have been recommended to be more physiological relevant, were not superior to the Transwell culture in either accuracy or convenience in drug permeability testing. Using rapid 3D printing prototyping techniques, this study proposed a hanging culture of Caco-2 cells that performed with high accuracy in predicting drug permeability in humans. As found, hanging cultured Caco-2 cells formed a confluent monolayer and maintained high cell viability on the 3D printed insert. Compared with the normal culture on Transwell, the Caco-2 cells on the 3D printed insert presented ∼30-100% higher brush border enzyme activity and ∼2-7 folds higher activity of P-glycoprotein/multidrug resistance-associated protein 2 during 21 days of incubation. For the eight membrane transporter substrates, the predictive curve of the 3D printing culture exhibited better linearity (R(2) = 0.92) to the human oral adsorption than that of the Transwell culture (R(2) = 0.84), indicating better prediction by the 3D printing culture. In this regard, the 3D printed insert for hanging culture could be potentially developed as a convenient and low-cost tool for testing drug oral absorption.

  2. Sample Preparation Strategies for Mass Spectrometry Imaging of 3D Cell Culture Models

    OpenAIRE

    Ahlf Wheatcraft, Dorothy R.; Liu, Xin; Hummon, Amanda B.

    2014-01-01

    Three dimensional cell cultures are attractive models for biological research. They combine the flexibility and cost-effectiveness of cell culture with some of the spatial and molecular complexity of tissue. For example, many cell lines form 3D structures given appropriate in vitro conditions. Colon cancer cell lines form 3D cell culture spheroids, in vitro mimics of avascular tumor nodules. While immunohistochemistry and other classical imaging methods are popular for monitoring the distribu...

  3. Development of 3-D Hydrogel Culture Systems With On-Demand Cell Separation

    OpenAIRE

    Hamilton, Sharon K.; Bloodworth, Nathaniel C.; Massad, Christopher S.; Hammoudi, Taymour M.; Suri, Shalu; Yang, Peter J.; Lu, Hang; Temenoff, Johnna S

    2013-01-01

    Recently there has been an increased interest in the effects of paracrine signaling between groups of cells, particularly in the context of better understanding how stem cells contribute to tissue repair. Most current 3-D co-culture methods lack the ability to effectively separate 2 cell populations after the culture period, which is important for simultaneously analyzing the reciprocal effects of each cell type on the other. Here, we detail the development of a 3-D hydrogel co-culture system...

  4. Printing Insecurity? The Security Implications of 3D-Printing of Weapons.

    Science.gov (United States)

    Walther, Gerald

    2015-12-01

    In 2013, the first gun printed out of plastic by a 3D-printer was successfully fired in the U.S. This event caused a major media hype about the dangers of being able to print a gun. Law enforcement agencies worldwide were concerned about this development and the potentially huge security implications of these functional plastic guns. As a result, politicians called for a ban of these weapons and a control of 3D-printing technology. This paper reviews the security implications of 3D-printing technology and 3D guns. It argues that current arms control and transfer policies are adequate to cover 3D-printed guns as well. However, while this analysis may hold up currently, progress in printing technology needs to be monitored to deal with future dangers pre-emptively.

  5. Bioimpedance monitoring of 3D cell culturing--complementary electrode configurations for enhanced spatial sensitivity.

    Science.gov (United States)

    Canali, Chiara; Heiskanen, Arto; Muhammad, Haseena Bashir; Høyum, Per; Pettersen, Fred-Johan; Hemmingsen, Mette; Wolff, Anders; Dufva, Martin; Martinsen, Ørjan Grøttem; Emnéus, Jenny

    2015-01-15

    A bioimpedance platform is presented as a promising tool for non-invasive real-time monitoring of the entire process of three-dimensional (3D) cell culturing in a hydrogel scaffold. In this study, the dynamics involved in the whole process of 3D cell culturing, starting from polymerisation of a bare 3D gelatin scaffold, to human mesenchymal stem cell (MSC) encapsulation and proliferation, was monitored over time. The platform consists of a large rectangular culture chamber with four embedded vertical gold plate electrodes that were exploited in two- and three terminal (2T and 3T) measurement configurations. By switching between the different combinations of electrode couples, it was possible to generate a multiplexing-like approach, which allowed for collecting spatially distributed information within the 3D space. Computational finite element (FE) analysis and electrochemical impedance spectroscopic (EIS) characterisation were used to determine the configurations' sensitivity field localisation. The 2T setup gives insight into the interfacial phenomena at both electrode surfaces and covers the central part of the 3D cell culture volume, while the four 3T modes provide focus on the dynamics at the corners of the 3D culture chamber. By combining a number of electrode configurations, complementary spatially distributed information on a large 3D cell culture can be obtained with maximised sensitivity in the entire 3D space. The experimental results show that cell proliferation can be monitored within the tested biomimetic environment, paving the way to further developments in bioimpedance tracking of 3D cell cultures and tissue engineering.

  6. Polysaccharide matrices used in 3D in vitro cell culture systems.

    Science.gov (United States)

    Diekjürgen, Dorina; Grainger, David W

    2017-10-01

    Polysaccharides comprise a diverse class of polymeric materials with a history of proven biocompatibility and continual use as biomaterials. Recent focus on new matrices appropriate for three-dimensional (3D) cell culture offers new opportunities to apply polysaccharides as extracellular matrix mimics. However, chemical and structural bases for specific cell-polysaccharide interactions essential for their utility as 3-D cell matrices are not well defined. This review describes how these naturally sourced biomaterials satisfy several key properties for current 3D cell culture needs and can also be synthetically modified or blended with additional components to tailor their cell engagement properties. Beyond their benign interactions with many cell types in cultures, their economical and high quality sourcing, optical clarity for ex situ analytical interrogation and in situ gelation represent important properties of these polymers for 3D cell culture applications. Continued diversification of their versatile glycan chemistry, new bio-synthetic sourcing strategies and elucidation of new cell-specific properties are attractive to expand the polysaccharide polymer utility for cell culture needs. Many 3D cell culture priorities are addressed with the portfolio of polysaccharide materials available and under development. This review provides a critical analysis of their properties, capabilities and challenges in 3D cell culture applications. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. A biofidelic 3D culture model to study the development of brain cellular systems

    Science.gov (United States)

    Ren, M.; Du, C.; Herrero Acero, E.; Tang-Schomer, M. D.; Özkucur, N.

    2016-01-01

    Little is known about how cells assemble as systems during corticogenesis to generate collective functions. We built a neurobiology platform that consists of fetal rat cerebral cortical cells grown within 3D silk scaffolds (SF). Ivermectin (Ivm), a glycine receptor (GLR) agonist, was used to modulate cell resting membrane potential (Vmem) according to methods described in a previous work that implicated Ivm in the arrangement and connectivity of cortical cell assemblies. The cells developed into distinct populations of neuroglial stem/progenitor cells, mature neurons or epithelial-mesenchymal cells. Importantly, the synchronized electrical activity in the newly developed cortical assemblies could be recorded as local field potential (LFP) measurements. This study therefore describes the first example of the development of a biologically relevant cortical plate assembly outside of the body. This model provides i) a preclinical basis for engineering cerebral cortex tissue autografts and ii) a biofidelic 3D culture model for investigating biologically relevant processes during the functional development of cerebral cortical cellular systems. PMID:27112667

  8. High-Quality 3d Models and Their Use in a Cultural Heritage Conservation Project

    Science.gov (United States)

    Tucci, G.; Bonora, V.; Conti, A.; Fiorini, L.

    2017-08-01

    Cultural heritage digitization and 3D modelling processes are mainly based on laser scanning and digital photogrammetry techniques to produce complete, detailed and photorealistic three-dimensional surveys: geometric as well as chromatic aspects, in turn testimony of materials, work techniques, state of preservation, etc., are documented using digitization processes. The paper explores the topic of 3D documentation for conservation purposes; it analyses how geomatics contributes in different steps of a restoration process and it presents an overview of different uses of 3D models for the conservation and enhancement of the cultural heritage. The paper reports on the project to digitize the earthenware frieze of the Ospedale del Ceppo in Pistoia (Italy) for 3D documentation, restoration work support, and digital and physical reconstruction and integration purposes. The intent to design an exhibition area suggests new ways to take advantage of 3D data originally acquired for documentation and scientific purposes.

  9. Bioimpedance monitoring of 3D cell culturing-Complementary electrode configurations for enhanced spatial sensitivity

    DEFF Research Database (Denmark)

    Canali, Chiara; Heiskanen, Arto; Muhammad, Haseena Bashir

    2015-01-01

    configurations. By switching between the different combinations of electrode couples, it was possible to generate a multiplexing-like approach, which allowed for collecting spatially distributed information within the 3D space. Computational finite element (FE) analysis and electrochemical impedance......A bioimpedance platform is presented as a promising tool for non-invasive real-time monitoring of the entire process of three-dimensional (3D) cell culturing in a hydrogel scaffold. In this study, the dynamics involved in the whole process of 3D cell culturing, starting from polymerisation...... of a bare 3D gelatin scaffold, to human mesenchymal stem cell (MSC) encapsulation and proliferation, was monitored over time. The platform consists of a large rectangular culture chamber with four embedded vertical gold plate electrodes that were exploited in two- and three terminal (2T and 3T) measurement...

  10. 3D cell culture to determine in vitro biocompatibility of bioactive glass in association with chitosan.

    Science.gov (United States)

    Bédouin, Y; Pellen Mussi, P; Tricot-Doleux, S; Chauvel-Lebret, D; Auroy, P; Ravalec, X; Oudadesse, H; Perez, F

    2015-01-01

    This study reports the in vitro biocompatibility of a composite biomaterial composed of 46S6 bioactive glass in association with chitosan (CH) by using 3D osteoblast culture of SaOS2. The 46S6 and CH composite (46S6-CH) forms small hydroxyapatite crystals on its surface after only three days immersion in the simulated body fluid. For 2D osteoblast culture, a significant increase in cell proliferation was observed after three days of contact with 46S6 or 46S6-CH-immersed media. After six days, 46S6-CH led to a significant increase in cell proliferation (128%) compared with pure 46S6 (113%) and pure CH (122%). For 3D osteoblast culture, after six days of culture, there was an increase in gene expression of markers of the early osteoblastic differentiation (RUNX2, ALP, COL1A1). Geometric structures corresponding to small apatite clusters were observed by SEM on the surface of the spheroids cultivated with 46S6 or 46S6-CH-immersed media. We showed different cellular responses depending on the 2D and 3D cell culture model. The induction of osteoblast differentiation in the 3D cell culture explained the differences of cell proliferation in contact with 46S6, CH or 46S6-CH-immersed media. This study confirmed that the 3D cell culture model is a very promising tool for in vitro biological evaluation of bone substitutes' properties.

  11. Peptide hydrogels – versatile matrices for 3D cell culture in cancer medicine

    Directory of Open Access Journals (Sweden)

    Peter eWorthington

    2015-04-01

    Full Text Available Traditional two-dimensional (2D cell culture systems have contributed tremendously to our understanding of cancer biology but have significant limitations in mimicking in vivo conditions such as the tumor microenvironment. In vitro, three-dimensional (3D cell culture models represent a more accurate, intermediate platform between simplified 2D culture models and complex and expensive in vivo models. 3D in vitro models can overcome 2D in vitro limitations caused by the oversupply of nutrients, and unphysiological cell-cell and cell-material interactions, and allow for dynamic interactions between cells, stroma, and extracellular matrix. In addition, 3D cultures allow for the development of concentration gradients, including oxygen, metabolites and growth factors, with chemical gradients playing an integral role in many cellular functions ranging from development to signaling in normal epithelia and cancer environments in vivo. Currently, the most common matrices used for 3D culture are biologically derived materials such as matrigel and collagen. However, in recent years, more defined, synthetic materials have become available as scaffolds for 3D culture with the advantage of forming well-defined, designed, tunable materials to control matrix charge, stiffness, porosity, nanostructure, degradability and adhesion properties, in addition to other material and biological properties. One important area of synthetic materials currently available for 3D cell culture are short sequence, self-assembling peptide hydrogels. In addition to the review of recent work towards the control of material, structure, and mechanical properties, we will also discuss the biochemical functionalization of peptide hydrogels and how this functionalization, coupled with desired hydrogel material characteristics, affects tumor cell behavior in 3D culture.

  12. Fabrication of 3D-culture platform with sandwich architecture for preserving liver-specific functions of hepatocytes using 3D bioprinter.

    Science.gov (United States)

    Arai, Kenichi; Yoshida, Toshiko; Okabe, Motonori; Goto, Mitsuaki; Mir, Tanveer Ahmad; Soko, Chika; Tsukamoto, Yoshinari; Akaike, Toshihiro; Nikaido, Toshio; Zhou, Kaixuan; Nakamura, Makoto

    2017-06-01

    The development of new three-dimensional (3D) cell culture system that maintains the physiologically relevant signals of hepatocytes is essential in drug discovery and tissue engineering research. Conventional two-dimensional (2D) culture yields cell growth, proliferation, and differentiation. However, gene expression and signaling profiles can be different from in vivo environment. Here, we report the fabrication of a 3D culture system using an artificial scaffold and our custom-made inkjet 3D bioprinter as a new strategy for studying liver-specific functions of hepatocytes. We built a 3D culture platform for hepatocytes-attachment and formation of cell monolayer by interacting the galactose chain of galactosylated alginate gel (GA-gel) with asialoglycoprotein receptor (ASGPR) of hepatocytes. The 3D geometrical arrangement of cells was controlled by using 3D bioprinter, and cell polarity was controlled with the galactosylated hydrogels. The fabricated GA-gel was able to successfully promote adhesion of hepatocytes. To observe liver-specific functions and to mimic hepatic cord, an additional parallel layer of hepatocytes was generated using two gel sheets. These results indicated that GA-gel biomimetic matrices can be used as a 3D culture system that could be effective for the engineering of liver tissues. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1583-1592, 2017. © 2017 Wiley Periodicals, Inc.

  13. Fabrication and optimization of alginate hydrogel constructs for use in 3D neural cell culture

    Energy Technology Data Exchange (ETDEWEB)

    Frampton, J P; Hynd, M R; Shain, W [Department of Biomedical Sciences, School of Public Health, State University of New York at Albany, Albany, NY 12210 (United States); Shuler, M L, E-mail: jf7674@albany.edu [Department of Biomedical Engineering, 270 Olin Hall, Cornell University, Ithaca, NY 14850 (United States)

    2011-02-15

    Two-dimensional (2D) culture systems provide useful information about many biological processes. However, some applications including tissue engineering, drug transport studies, and analysis of cell growth and dynamics are better studied using three-dimensional (3D) culture systems. 3D culture systems can potentially offer higher degrees of organization and control of cell growth environments, more physiologically relevant diffusion characteristics, and permit the formation of more extensive 3D networks of cell-cell interactions. A 3D culture system has been developed using alginate as a cell scaffold, capable of maintaining the viability and function of a variety of neural cell types. Alginate was functionalized by the covalent attachment of a variety of whole proteins and peptide epitopes selected to provide sites for cell attachment. Alginate constructs were used to entrap a variety of neural cell types including astroglioma cells, astrocytes, microglia and neurons. Neural cells displayed process outgrowth over time in culture. Cell-seeded scaffolds were characterized in terms of their biochemical and biomechanical properties, effects on seeded neural cells, and suitability for use as 3D neural cell culture models.

  14. Effects of 3D microwell culture on initial fate specification in human embryonic stem cells.

    Science.gov (United States)

    Hsiao, Cheston; Tomai, Matthew; Glynn, Jeremy; Palecek, Sean P

    2014-04-01

    Several studies have demonstrated that 3D culture systems influence human embryonic stem cell (hESC) phenotypes and fate choices. However, the effect that these microenvironmental changes have on signaling pathways governing hESC behaviors is not well understood. Here, we have used a 3D microwell array to investigate differences in activation of developmental pathways between 2D and 3D cultures of both undifferentiated hESCs and hESCs undergoing initial differentiation in embryoid bodies (EBs). We observed increased induction into mesoderm and endoderm and differences in expression of genes from multiple signaling pathways that regulate development, including Wnt/β-catenin, TGF-β superfamily, Notch and FGF during EB-mediated differentiation, in 3D microwells as compared with the 2D substrates. In undifferentiated hESCs, we also observed differences in epithelial-mesenchymal transition phenotypes and the TGFβ/BMP pathway between cultures in 3D and 2D. These results illustrate that 3D culture influences multiple pathways that may regulate the differentiation trajectories of hESCs.

  15. INTEGRATED DATA CAPTURING REQUIREMENTS FOR 3D SEMANTIC MODELLING OF CULTURAL HERITAGE: THE INCEPTION PROTOCOL

    Directory of Open Access Journals (Sweden)

    R. Di Giulio

    2017-02-01

    In order to face these challenges and to start solving the issue of the large amount of captured data and time-consuming processes in the production of 3D digital models, an Optimized Data Acquisition Protocol (DAP has been set up. The purpose is to guide the processes of digitization of cultural heritage, respecting needs, requirements and specificities of cultural assets.

  16. Nanoparticle toxicity assessment using an in vitro 3-D kidney organoid culture model.

    Science.gov (United States)

    Astashkina, Anna I; Jones, Clint F; Thiagarajan, Giridhar; Kurtzeborn, Kristen; Ghandehari, Hamid; Brooks, Benjamin D; Grainger, David W

    2014-08-01

    Nanocarriers and nanoparticles remain an intense pharmaceutical and medical imaging technology interest. Their entry into clinical use is hampered by the lack of reliable in vitro models that accurately predict in vivo toxicity. This study evaluates a 3-D kidney organoid proximal tubule culture to assess in vitro toxicity of the hydroxylated generation-5 PAMAM dendrimer (G5-OH) compared to previously published preclinical in vivo rodent nephrotoxicity data. 3-D kidney proximal tubule cultures were created using isolated murine proximal tubule fractions suspended in a biomedical grade hyaluronic acid-based hydrogel. Toxicity in these cultures to neutral G5-OH dendrimer nanoparticles and gold nanoparticles in vitro was assessed using clinical biomarker generation. Neutral PAMAM nanoparticle dendrimers elicit in vivo-relevant kidney biomarkers and cell viability in a 3-D kidney organoid culture that closely reflect toxicity markers reported in vivo in rodent nephrotoxicity models exposed to this same nanoparticle.

  17. In vivo biomarker expression patterns are preserved in 3D cultures of Prostate Cancer

    Energy Technology Data Exchange (ETDEWEB)

    Windus, Louisa C.E.; Kiss, Debra L.; Glover, Tristan [Eskitis Institute for Cell and Molecular Therapies, Discovery Biology, Griffith University, Nathan 4111, Brisbane, Queensland (Australia); Avery, Vicky M., E-mail: v.avery@griffith.edu.au [Eskitis Institute for Cell and Molecular Therapies, Discovery Biology, Griffith University, Nathan 4111, Brisbane, Queensland (Australia)

    2012-11-15

    Here we report that Prostate Cancer (PCa) cell-lines DU145, PC3, LNCaP and RWPE-1 grown in 3D matrices in contrast to conventional 2D monolayers, display distinct differences in cell morphology, proliferation and expression of important biomarker proteins associated with cancer progression. Consistent with in vivo growth rates, in 3D cultures, all PCa cell-lines were found to proliferate at significantly lower rates in comparison to their 2D counterparts. Moreover, when grown in a 3D matrix, metastatic PC3 cell-lines were found to mimic more precisely protein expression patterns of metastatic tumour formation as found in vivo. In comparison to the prostate epithelial cell-line RWPE-1, metastatic PC3 cell-lines exhibited a down-regulation of E-cadherin and {alpha}6 integrin expression and an up-regulation of N-cadherin, Vimentin and {beta}1 integrin expression and re-expressed non-transcriptionally active AR. In comparison to the non-invasive LNCaP cell-lines, PC3 cells were found to have an up-regulation of chemokine receptor CXCR4, consistent with a metastatic phenotype. In 2D cultures, there was little distinction in protein expression between metastatic, non-invasive and epithelial cells. These results suggest that 3D cultures are more representative of in vivo morphology and may serve as a more biologically relevant model in the drug discovery pipeline. -- Highlights: Black-Right-Pointing-Pointer We developed and optimised 3D culturing techniques for Prostate Cancer cell-lines. Black-Right-Pointing-Pointer We investigated biomarker expression in 2D versus 3D culture techniques. Black-Right-Pointing-Pointer Metastatic PC3 cells re-expressed non-transcriptionally active androgen receptor. Black-Right-Pointing-Pointer Metastatic PCa cell lines retain in vivo-like antigenic profiles in 3D cultures.

  18. Application of 3D printing to prototype and develop novel plant tissue culture systems.

    Science.gov (United States)

    Shukla, Mukund R; Singh, Amritpal S; Piunno, Kevin; Saxena, Praveen K; Jones, A Maxwell P

    2017-01-01

    Due to the complex process of designing and manufacturing new plant tissue culture vessels through conventional means there have been limited efforts to innovate improved designs. Further, development and availability of low cost, energy efficient LEDs of various spectra has made it a promising light source for plant growth in controlled environments. However, direct replacement of conventional lighting sources with LEDs does not address problems with uniformity, spectral control, or the challenges in conducting statistically valid experiments to assess the effects of light. Prototyping using 3D printing and LED based light sources could help overcome these limitations and lead to improved culture systems. A modular culture vessel design in which the fluence rate and spectrum of light are independently controlled was designed, prototyped using 3D printing, and evaluated for plant growth. This design is compatible with semi-solid and liquid based culture systems. Observations on morphology, chlorophyll content, and chlorophyll fluorescence based stress parameters from in vitro plants cultured under different light spectra with similar overall fluence rate indicated different responses in Nicotiana tabacum and Artemisia annua plantlets. This experiment validates the utility of 3D printing to design and test functional vessels and demonstrated that optimal light spectra for in vitro plant growth is species-specific. 3D printing was successfully used to prototype novel culture vessels with independently controlled variable fluence rate/spectra LED lighting. This system addresses several limitations associated with current lighting systems, providing more uniform lighting and allowing proper replication/randomization for experimental plant biology while increasing energy efficiency. A complete procedure including the design and prototyping of a culture vessel using 3D printing, commercial scale injection molding of the prototype, and conducting a properly replicated

  19. 3D culture broadly regulates tumor cell hypoxia response and angiogenesis via pro-inflammatory pathways.

    Science.gov (United States)

    DelNero, Peter; Lane, Maureen; Verbridge, Scott S; Kwee, Brian; Kermani, Pouneh; Hempstead, Barbara; Stroock, Abraham; Fischbach, Claudia

    2015-07-01

    Oxygen status and tissue dimensionality are critical determinants of tumor angiogenesis, a hallmark of cancer and an enduring target for therapeutic intervention. However, it is unclear how these microenvironmental conditions interact to promote neovascularization, due in part to a lack of comprehensive, unbiased data sets describing tumor cell gene expression as a function of oxygen levels within three-dimensional (3D) culture. Here, we utilized alginate-based, oxygen-controlled 3D tumor models to study the interdependence of culture context and the hypoxia response. Microarray gene expression analysis of tumor cells cultured in 2D versus 3D under ambient or hypoxic conditions revealed striking interdependence between culture dimensionality and hypoxia response, which was mediated in part by pro-inflammatory signaling pathways. In particular, interleukin-8 (IL-8) emerged as a major player in the microenvironmental regulation of the hypoxia program. Notably, this interaction between dimensionality and oxygen status via IL-8 increased angiogenic sprouting in a 3D endothelial invasion assay. Taken together, our data suggest that pro-inflammatory pathways are critical regulators of tumor hypoxia response within 3D environments that ultimately impact tumor angiogenesis, potentially providing important therapeutic targets. Furthermore, these results highlight the importance of pathologically relevant tissue culture models to study the complex physical and chemical processes by which the cancer microenvironment mediates new vessel formation.

  20. 3D printing – a key technology for tailored biomedical cell culture lab ware

    Directory of Open Access Journals (Sweden)

    Schmieder Florian

    2016-09-01

    Full Text Available Today’s 3D printing technologies offer great possibilities for biomedical researchers to create their own specific laboratory equipment. With respect to the generation of ex vivo vascular perfusion systems this will enable new types of products that will embed complex 3D structures possibly coupled with cell loaded scaffolds closely reflecting the in-vivo environment. Moreover this could lead to microfluidic devices that should be available in small numbers of pieces at moderate prices. Here, we will present first results of such 3D printed cell culture systems made from plastics and show their use for scaffold based applications.

  1. 3D Modeling from Multi-views Images for Cultural Heritage in Wat-Pho, Thailand

    OpenAIRE

    N. Soontranon; Srestasathiern, P.; Lawawirojwong, S.

    2015-01-01

    In Thailand, there are several types of (tangible) cultural heritages. This work focuses on 3D modeling of the heritage objects from multi-views images. The images are acquired by using a DSLR camera which costs around $1,500 (camera and lens). Comparing with a 3D laser scanner, the camera is cheaper and lighter than the 3D scanner. Hence, the camera is available for public users and convenient for accessing narrow areas. The acquired images consist of various sculptures and architec...

  2. 3D Modeling from Multi-views Images for Cultural Heritage in Wat-Pho, Thailand

    Science.gov (United States)

    Soontranon, N.; Srestasathiern, P.; Lawawirojwong, S.

    2015-08-01

    In Thailand, there are several types of (tangible) cultural heritages. This work focuses on 3D modeling of the heritage objects from multi-views images. The images are acquired by using a DSLR camera which costs around 1,500 (camera and lens). Comparing with a 3D laser scanner, the camera is cheaper and lighter than the 3D scanner. Hence, the camera is available for public users and convenient for accessing narrow areas. The acquired images consist of various sculptures and architectures in Wat-Pho which is a Buddhist temple located behind the Grand Palace (Bangkok, Thailand). Wat-Pho is known as temple of the reclining Buddha and the birthplace of traditional Thai massage. To compute the 3D models, a diagram is separated into following steps; Data acquisition, Image matching, Image calibration and orientation, Dense matching and Point cloud processing. For the initial work, small heritages less than 3 meters height are considered for the experimental results. A set of multi-views images of an interested object is used as input data for 3D modeling. In our experiments, 3D models are obtained from MICMAC (open source) software developed by IGN, France. The output of 3D models will be represented by using standard formats of 3D point clouds and triangulated surfaces such as .ply, .off, .obj, etc. To compute for the efficient 3D models, post-processing techniques are required for the final results e.g. noise reduction, surface simplification and reconstruction. The reconstructed 3D models can be provided for public access such as website, DVD, printed materials. The high accurate 3D models can also be used as reference data of the heritage objects that must be restored due to deterioration of a lifetime, natural disasters, etc.

  3. Microchip-based 3D-Cell Culture Using Polymer Nanofibers Generated by Solution Blow Spinning.

    Science.gov (United States)

    Chen, Chengpeng; Townsend, Alexandra D; Sell, Scott A; Martin, R Scott

    2017-06-14

    Polymer nano/micro fibers have found many applications including 3D cell culture and the creation of wound dressings. The fibers can be produced by a variety of techniques that include electrospinning, the primary disadvantage of which include the requirement for a high voltage supply (which may cause issues such as polymer denaturation) and lack of portability. More recently, solution blow spinning, where a high velocity sheath gas is used instead of high voltage, has been used to generate polymer fibers. In this work, we used blow spinning to create nano/microfibers for microchip-based 3D cell culture. First, we thoroughly investigated fiber generation from a 3D printed gas sheath device using two polymers that are amenable to cell culture (polycaprolactone, PCL and polystyrene, PS) as well as the parameters that can affect PCL and PS fiber quality. Using the 3D printed sheath device, it was found that the pressure of the sheath N2 and the concentration of polymer solutions determine if fibers can be produced as well as the resulting fiber morphology. In addition, we showed how these fibers can be used for 3D cell culture by directly depositing PCL fibers in petri dishes and well plates. It is shown the fibers have good compatibility with RAW 264.7 macrophages and the PCL fiber scaffold can be as thick as 178 ± 14 μm. PCL fibers created from solution blow spinning (with the 3D printed sheath device) were then integrated with a microfluidic device for the first time to fabricate a 3D cell culture scaffold with a flow component. After culturing and stimulating macrophages on the fluidic device, it was found that the integrated 3D fibrous scaffold is a better mimic of the extracellular matrix (as opposed to a flat, 2D substrate), with enhanced nitrite accumulation (product of nitric oxide release) from macrophages stimulated with lipopolysaccharide. PS fibers were also made and integrated in a microfluidic device for 3D culture of endothelial cells, which stayed

  4. Modeling spatial distribution of oxygen in 3d culture of islet beta-cells.

    Science.gov (United States)

    McReynolds, John; Wen, Yu; Li, Xiaofei; Guan, Jianjun; Jin, Sha

    2017-01-01

    Three-dimensional (3D) scaffold culture of pancreatic β-cell has been proven to be able to better mimic physiological conditions in the body. However, one critical issue with culturing pancreatic β-cells is that β-cells consume large amounts of oxygen, and hence insufficient oxygen supply in the culture leads to loss of β-cell mass and functions. This becomes more significant when cells are cultured in a 3D scaffold. In this study, in order to understand the effect of oxygen tension inside a cell-laden collagen culture on β-cell proliferation, a culture model with encapsulation of an oxygen-generator was established. The oxygen-generator was made by embedding hydrogen peroxide into nontoxic polydimethylsiloxane to avoid the toxicity of a chemical reaction in the β-cell culture. To examine the effectiveness of the oxygenation enabled 3D culture, the spatial-temporal distribution of oxygen tension inside a scaffold was evaluated by a mathematical modeling approach. Our simulation results indicated that an oxygenation-aided 3D culture would augment the oxygen supply required for the β-cells. Furthermore, we identified that cell seeding density and the capacity of the oxygenator are two critical parameters in the optimization of the culture. Notably, cell-laden scaffold cultures with an in situ oxygen supply significantly improved the β-cells' biological function. These β-cells possess high insulin secretion capacity. The results obtained in this work would provide valuable information for optimizing and encouraging functional β-cell cultures. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 33:221-228, 2017.

  5. Using a 3D Culture System to Differentiate Visceral Adipocytes In Vitro.

    Science.gov (United States)

    Emont, Margo P; Yu, Hui; Jun, Heejin; Hong, Xiaowei; Maganti, Nenita; Stegemann, Jan P; Wu, Jun

    2015-12-01

    It has long been recognized that body fat distribution and regional adiposity play a major role in the control of metabolic homeostasis. However, the ability to study and compare the cell autonomous regulation and response of adipocytes from different fat depots has been hampered by the difficulty of inducing preadipocytes isolated from the visceral depot to differentiate into mature adipocytes in culture. Here, we present an easily created 3-dimensional (3D) culture system that can be used to differentiate preadipocytes from the visceral depot as robustly as those from the sc depot. The cells differentiated in these 3D collagen gels are mature adipocytes that retain depot-specific characteristics, as determined by imaging, gene expression, and functional assays. This 3D culture system therefore allows for study of the development and function of adipocytes from both depots in vitro and may ultimately lead to a greater understanding of site-specific functional differences of adipose tissues to metabolic dysregulation.

  6. Controlled 3D culture in Matrigel microbeads to analyze clonal acinar development.

    Science.gov (United States)

    Dolega, Monika E; Abeille, Fabien; Picollet-D'hahan, Nathalie; Gidrol, Xavier

    2015-06-01

    3D culture systems are a valuable tool for modeling morphogenesis and carcinogenesis of epithelial tissue in a structurally appropriate context. We present a novel approach for 3D cell culture based on a flow-focusing microfluidic system that encapsulates epithelial cells in Matrigel beads. As a model we use prostatic and breast cells and assay for development of acini, polarized cellular spheres enclosing lumen. Each individual bead on average acts as a single 3D cell culture compartment generating one acinus per bead. Compared to standard protocols microfluidics provides increased control over the environment leading to more a uniform acini population. The increased facility of bead manipulation allowed us to isolate single cells which are self-sufficient to fully develop into acini in presence of Matrigel. Furthermore, combination of our microfluidic approach with large particle FACS opens new avenues in high throughput screening on single acini or spheroids.

  7. Imaging of Metabolic Status in 3D Cultures with an Improved AMPK FRET Biosensor for FLIM

    Directory of Open Access Journals (Sweden)

    George Chennell

    2016-08-01

    Full Text Available We describe an approach to non-invasively map spatiotemporal biochemical and physiological changes in 3D cell culture using Forster Resonance Energy Transfer (FRET biosensors expressed in tumour spheroids. In particular, we present an improved Adenosine Monophosphate (AMP Activated Protein Kinase (AMPK FRET biosensor, mTurquoise2 AMPK Activity Reporter (T2AMPKAR, for fluorescence lifetime imaging (FLIM readouts that we have evaluated in 2D and 3D cultures. Our results in 2D cell culture indicate that replacing the FRET donor, enhanced Cyan Fluorescent Protein (ECFP, in the original FRET biosensor, AMPK activity reporter (AMPKAR, with mTurquoise2 (mTq2FP, increases the dynamic range of the response to activation of AMPK, as demonstrated using the direct AMPK activator, 991. We demonstrated 3D FLIM of this T2AMPKAR FRET biosensor expressed in tumour spheroids using two-photon excitation.

  8. User-Appropriate Viewer for High Resolution Interactive Engagement with 3d Digital Cultural Artefacts

    Science.gov (United States)

    Gillespie, D.; La Pensée, A.; Cooper, M.

    2013-07-01

    Three dimensional (3D) laser scanning is an important documentation technique for cultural heritage. This technology has been adopted from the engineering and aeronautical industry and is an invaluable tool for the documentation of objects within museum collections (La Pensée, 2008). The datasets created via close range laser scanning are extremely accurate and the created 3D dataset allows for a more detailed analysis in comparison to other documentation technologies such as photography. The dataset can be used for a range of different applications including: documentation; archiving; surface monitoring; replication; gallery interactives; educational sessions; conservation and visualization. However, the novel nature of a 3D dataset is presenting a rather unique challenge with respect to its sharing and dissemination. This is in part due to the need for specialised 3D software and a supported graphics card to display high resolution 3D models. This can be detrimental to one of the main goals of cultural institutions, which is to share knowledge and enable activities such as research, education and entertainment. This has limited the presentation of 3D models of cultural heritage objects to mainly either images or videos. Yet with recent developments in computer graphics, increased internet speed and emerging technologies such as Adobe's Stage 3D (Adobe, 2013) and WebGL (Khronos, 2013), it is now possible to share a dataset directly within a webpage. This allows website visitors to interact with the 3D dataset allowing them to explore every angle of the object, gaining an insight into its shape and nature. This can be very important considering that it is difficult to offer the same level of understanding of the object through the use of traditional mediums such as photographs and videos. Yet this presents a range of problems: this is a very novel experience and very few people have engaged with 3D objects outside of 3D software packages or games. This paper

  9. IMAGO Visualization System: An Interactive Web-Based 3D Visualization System for Cultural Heritage Applications

    Directory of Open Access Journals (Sweden)

    Caroline M. Mendes

    2012-04-01

    Full Text Available Due to the evolution of technologies and methods for realistic 3D reconstruction of objects, in many projects it can be found efficient ways to make research results in digital preservation available on the Internet. 3D visualization of cultural heritage is highlighted in this scenario, helping to expand research activities in this field by providing proper tools to allow for example, remote access to historical artifacts. Thus, visualization systems must be able to handle important aspects in the context of digital preservation, such as user profiles, security and ease access to 3D models. This paper presents the development of an effective web-based 3D visualization system whose architecture offers an easy and fast interactivity with 3D models even when limited computer resources are available. The system has been successfully adopted in developing of 3D Virtual Museums in the Universidade Federal do Parana (UFPR in Brazil, providing an important tool to promote research, educational, social and cultural activities.

  10. BioSig3D: High Content Screening of Three-Dimensional Cell Culture Models.

    Directory of Open Access Journals (Sweden)

    Cemal Cagatay Bilgin

    Full Text Available BioSig3D is a computational platform for high-content screening of three-dimensional (3D cell culture models that are imaged in full 3D volume. It provides an end-to-end solution for designing high content screening assays, based on colony organization that is derived from segmentation of nuclei in each colony. BioSig3D also enables visualization of raw and processed 3D volumetric data for quality control, and integrates advanced bioinformatics analysis. The system consists of multiple computational and annotation modules that are coupled together with a strong use of controlled vocabularies to reduce ambiguities between different users. It is a web-based system that allows users to: design an experiment by defining experimental variables, upload a large set of volumetric images into the system, analyze and visualize the dataset, and either display computed indices as a heatmap, or phenotypic subtypes for heterogeneity analysis, or download computed indices for statistical analysis or integrative biology. BioSig3D has been used to profile baseline colony formations with two experiments: (i morphogenesis of a panel of human mammary epithelial cell lines (HMEC, and (ii heterogeneity in colony formation using an immortalized non-transformed cell line. These experiments reveal intrinsic growth properties of well-characterized cell lines that are routinely used for biological studies. BioSig3D is being released with seed datasets and video-based documentation.

  11. Metadata and Tools for Integration and Preservation of Cultural Heritage 3D Information

    Directory of Open Access Journals (Sweden)

    Achille Felicetti

    2011-12-01

    Full Text Available In this paper we investigate many of the various storage, portability and interoperability issues arising among archaeologists and cultural heritage people when dealing with 3D technologies. On the one side, the available digital repositories look often unable to guarantee affordable features in the management of 3D models and their metadata; on the other side the nature of most of the available data format for 3D encoding seem to be not satisfactory for the necessary portability required nowadays by 3D information across different systems. We propose a set of possible solutions to show how integration can be achieved through the use of well known and wide accepted standards for data encoding and data storage. Using a set of 3D models acquired during various archaeological campaigns and a number of open source tools, we have implemented a straightforward encoding process to generate meaningful semantic data and metadata. We will also present the interoperability process carried out to integrate the encoded 3D models and the geographic features produced by the archaeologists. Finally we will report the preliminary (rather encouraging development of a semantic enabled and persistent digital repository, where 3D models (but also any kind of digital data and metadata can easily be stored, retrieved and shared with the content of other digital archives.

  12. BioSig3D: High Content Screening of Three-Dimensional Cell Culture Models.

    Science.gov (United States)

    Bilgin, Cemal Cagatay; Fontenay, Gerald; Cheng, Qingsu; Chang, Hang; Han, Ju; Parvin, Bahram

    2016-01-01

    BioSig3D is a computational platform for high-content screening of three-dimensional (3D) cell culture models that are imaged in full 3D volume. It provides an end-to-end solution for designing high content screening assays, based on colony organization that is derived from segmentation of nuclei in each colony. BioSig3D also enables visualization of raw and processed 3D volumetric data for quality control, and integrates advanced bioinformatics analysis. The system consists of multiple computational and annotation modules that are coupled together with a strong use of controlled vocabularies to reduce ambiguities between different users. It is a web-based system that allows users to: design an experiment by defining experimental variables, upload a large set of volumetric images into the system, analyze and visualize the dataset, and either display computed indices as a heatmap, or phenotypic subtypes for heterogeneity analysis, or download computed indices for statistical analysis or integrative biology. BioSig3D has been used to profile baseline colony formations with two experiments: (i) morphogenesis of a panel of human mammary epithelial cell lines (HMEC), and (ii) heterogeneity in colony formation using an immortalized non-transformed cell line. These experiments reveal intrinsic growth properties of well-characterized cell lines that are routinely used for biological studies. BioSig3D is being released with seed datasets and video-based documentation.

  13. A 3D modeling and measurement system for cultural heritage preservation

    Science.gov (United States)

    Du, Guoguang; Zhou, Mingquan; Ren, Pu; Shui, Wuyang; Zhou, Pengbo; Wu, Zhongke

    2015-07-01

    Cultural Heritage reflects the human production, life style and environmental conditions of various historical periods. It exists as one of the major national carriers of national history and culture. In order to do better protection and utilization for these cultural heritages, a system of three-dimensional (3D) reconstruction and statistical measurement is proposed in this paper. The system solves the problems of cultural heritage's data storage, measurement and analysis. Firstly, for the high precision modeling and measurement problems, range data registration and integration algorithm used to achieve high precision 3D reconstruction. Secondly, multi-view stereo reconstruction method is used to solve the problem of rapid reconstruction by procedures such as the original image data pre-processing, camera calibration, point cloud modeling. At last, the artifacts' measure underlying database is established by calculating the measurements of the 3D model's surface. These measurements contain Euclidean distance between the points on the surface, geodesic distance between the points, normal and curvature in each point, superficial area of a region, volume of model's part and some other measurements. These measurements provide a basis for carrying out information mining of cultural heritage. The system has been applied to the applications of 3D modeling, data measurement of the Terracotta Warriors relics, Tibetan architecture and some other relics.

  14. 3D cell culture: a review of current approaches and techniques.

    Science.gov (United States)

    Haycock, John W

    2011-01-01

    Cell culture in two dimensions has been routinely and diligently undertaken in thousands of laboratories worldwide for the past four decades. However, the culture of cells in two dimensions is arguably primitive and does not reproduce the anatomy or physiology of a tissue for informative or useful study. Creating a third dimension for cell culture is clearly more relevant, but requires a multidisciplinary approach and multidisciplinary expertise. When entering the third dimension, investigators need to consider the design of scaffolds for supporting the organisation of cells or the use of bioreactors for controlling nutrient and waste product exchange. As 3D culture systems become more mature and relevant to human and animal physiology, the ability to design and develop co-cultures becomes possible as does the ability to integrate stem cells. The primary objectives for developing 3D cell culture systems vary widely - and range from engineering tissues for clinical delivery through to the development of models for drug screening. The intention of this review is to provide a general overview of the common approaches and techniques for designing 3D culture models.

  15. 3D culture increases pluripotent gene expression in mesenchymal stem cells through relaxation of cytoskeleton tension.

    Science.gov (United States)

    Zhou, Ying; Chen, Haiyan; Li, Hong; Wu, Yaojiong

    2017-03-09

    Three-dimensional (3D) culture has been shown to improve pluripotent gene expression in mesenchymal stem cells (MSCs), but the underlining mechanisms were poorly understood. Here, we found that the relaxation of cytoskeleton tension of MSCs in 3D culture was critically associated with the expressional up-regulation of Nanog. Cultured in spheroids, MSCs showed decreased integrin-based cell-matrix adhesion but increased cadherin-based cell-cell interaction. Different from that in 2D culture, where MSCs exhibited branched and multiple-directed F-actin stress bundles at the cell edge and strengthened stress fibres transversing the cell body, MSCs cultured in spheroids showed compact cell body, relaxed cytoskeleton tension with very thin cortical actin filament outlining the cell, and increased expression of Nanog along with reduced levels of Suv39h1 (H3K9 methyltransferase) and H3K9me3. Notably, pharmaceutical inhibition of actin polymerization with cytochalasin D or silencing Suv39h1 expression with siRNA in 2D-cultured MSCs elevated the expression of Nanog via H3K9 demethylation. Thus, our data suggest that 3D culture increases the expression of Nanog through the relaxation of actin cytoskeleton, which mediates reduced Suv39h1 and H3K9me3 levels.

  16. Functional 3-D cardiac co-culture model using bioactive chitosan nanofiber scaffolds.

    Science.gov (United States)

    Hussain, Ali; Collins, George; Yip, Derek; Cho, Cheul H

    2013-02-01

    The in vitro generation of a three-dimensional (3-D) myocardial tissue-like construct employing cells, biomaterials, and biomolecules is a promising strategy in cardiac tissue regeneration, drug testing, and tissue engineering applications. Despite significant progress in this field, current cardiac tissue models are not yet able to stably maintain functional characteristics of cardiomyocytes for long-term culture and therapeutic purposes. The objective of this study was to fabricate bioactive 3-D chitosan nanofiber scaffolds using an electrospinning technique and exploring its potential for long-term cardiac function in the 3-D co-culture model. Chitosan is a natural polysaccharide biomaterial that is biocompatible, biodegradable, non-toxic, and cost effective. Electrospun chitosan was utilized to provide structural scaffolding characterized by scale and architectural resemblance to the extracellular matrix (ECM) in vivo. The chitosan fibers were coated with fibronectin via adsorption in order to enhance cellular adhesion to the fibers and migration into the interfibrous milieu. Ventricular cardiomyocytes were harvested from neonatal rats and studied in various culture conditions (i.e., mono- and co-cultures) for their viability and function. Cellular morphology and functionality were examined using immunofluorescent staining for alpha-sarcomeric actin (SM-actin) and gap junction protein, Connexin-43 (Cx43). Scanning electron microscopy (SEM) and light microscopy were used to investigate cellular morphology, spatial organization, and contractions. Calcium indicator was used to monitor calcium ion flux of beating cardiomyocytes. The results demonstrate that the chitosan nanofibers retained their cylindrical morphology in long-term cell cultures and exhibited good cellular attachment and spreading in the presence of adhesion molecule, fibronectin. Cardiomyocyte mono-cultures resulted in loss of cardiomyocyte polarity and islands of non-coherent contractions. However

  17. Nuclear Factor-kappaB controls the reaggregation of 3D neurosphere cultures in vitro

    Directory of Open Access Journals (Sweden)

    D Widera

    2006-05-01

    Full Text Available The approach of reaggregation involves the regeneration and self-renewal of histotypical 3D spheres from isolated tissue kept in suspension culture. Reaggregated spheres can be used as tumour, genetic, biohybrid and neurosphere models. In addition the functional superiority of 3D aggregates over conventional 2D cultures developed the use of neurospheres for brain engineering of CNS diseases. Thus 3D aggregate cultures created enormous interest in mechanisms that regulate the formation of multicellular aggregates in vitro. Here we analyzed mechanisms guiding the development of 3D neurosphere cultures. Adult neural stem cells can be cultured as self-adherent clusters, called neurospheres. Neurospheres are characterised as heterogeneous clusters containing unequal stem cell sub-types. Tumour necrosis factor-alpha (TNF- alpha is one of the crucial inflammatory cytokines with multiple actions on several cell types. TNF- alpha strongly activates the canonical Nuclear Factor Kappa-B (NF-kappaB pathway. In order to investigate further functions of TNF in neural stem cells (NSCs we tested the hypothesis that TNF is able to modulate the motility and/or migratory behaviour of SVZ derived adult neural stem cells. We observed a significantly faster sphere formation in TNF treated cultures than in untreated controls. The very fast aggregation of isolated NSCs (<2h is a commonly observed phenomenon, though the mechanisms of 3D neurosphere formation remain largely unclear. Here we demonstrate for the first time, increased aggregation and enhanced motility of isolated NSCs in response to the TNF-stimulus. Moreover, this phenomenon is largely dependent on activated transcription factor NF-kappaB. Both, the pharmacological blockade of NF-kappaB pathway by pyrrolidine dithiocarbamate (PDTC or Bay11-7082 and genetic blockade by expression of a transdominant-negative super-repressor IkappaB-AA1 led to decreased aggregation.

  18. Recovery of important physiological functions in 3D culture of immortal hepatocytes

    DEFF Research Database (Denmark)

    Wrzesinski, Krzysztof; Fey, S. J.

    2011-01-01

    hepatocytes using the classical approaches (in “2D”) and using a system which leads to the generation of spheroids of cells held in suspension (“3D”). Both approaches gave rise to cultures where the large majority of cells were viable, produced similar amounts of ATP, incorporated similar amounts......It is widely expected that cells grown in 3D environments (in suspension, on scaffolds etc.) will be superior to growing cells in classical 2D culture flasks. These expectations include the belief that cells grown in 3D culture will possess physiological characteristics that resemble more closely...... to grow human liver cells in ‘3 dimensional’ cultures so that they behave very similar to the liver in our bodies. By growing the immortal hepatocytes in specially designed bioreactors they form small pieces of ‘pseudotissue’ which exhibit several of the functions seen in the adult liver. We have grown...

  19. 3D Culture as a Clinically Relevant Model for Personalized Medicine.

    Science.gov (United States)

    Fong, Eliza Li Shan; Toh, Tan Boon; Yu, Hanry; Chow, Edward Kai-Hua

    2017-03-01

    Advances in understanding many of the fundamental mechanisms of cancer progression have led to the development of molecular targeted therapies. While molecular targeted therapeutics continue to improve the outcome for cancer patients, tumor heterogeneity among patients, as well as intratumoral heterogeneity, limits the efficacy of these drugs to specific patient subtypes, as well as contributes to relapse. Thus, there is a need for a more personalized approach toward drug development and diagnosis that takes into account the diversity of cancer patients, as well as the complex milieu of tumor cells within a single patient. Three-dimensional (3D) culture systems paired with patient-derived xenografts or patient-derived organoids may provide a more clinically relevant system to address issues presented by personalized or precision medical approaches. In this review, we cover the current methods available for applying 3D culture systems toward personalized cancer research and drug development, as well as key challenges that must be addressed in order to fully realize the potential of 3D patient-derived culture systems for cancer drug development. Greater implementation of 3D patient-derived culture systems in the cancer research field should accelerate the development of truly personalized medical therapies for cancer patients.

  20. Polymer-based mesh as supports for multi-layered 3D cell culture and assays.

    Science.gov (United States)

    Simon, Karen A; Park, Kyeng Min; Mosadegh, Bobak; Subramaniam, Anand Bala; Mazzeo, Aaron D; Ngo, Philip M; Whitesides, George M

    2014-01-01

    Three-dimensional (3D) culture systems can mimic certain aspects of the cellular microenvironment found in vivo, but generation, analysis and imaging of current model systems for 3D cellular constructs and tissues remain challenging. This work demonstrates a 3D culture system-Cells-in-Gels-in-Mesh (CiGiM)-that uses stacked sheets of polymer-based mesh to support cells embedded in gels to form tissue-like constructs; the stacked sheets can be disassembled by peeling the sheets apart to analyze cultured cells-layer-by-layer-within the construct. The mesh sheets leave openings large enough for light to pass through with minimal scattering, and thus allowing multiple options for analysis-(i) using straightforward analysis by optical light microscopy, (ii) by high-resolution analysis with fluorescence microscopy, or (iii) with a fluorescence gel scanner. The sheets can be patterned into separate zones with paraffin film-based decals, in order to conduct multiple experiments in parallel; the paraffin-based decal films also block lateral diffusion of oxygen effectively. CiGiM simplifies the generation and analysis of 3D culture without compromising throughput, and quality of the data collected: it is especially useful in experiments that require control of oxygen levels, and isolation of adjacent wells in a multi-zone format.

  1. Bioengineered 3D Glial Cell Culture Systems and Applications for Neurodegeneration and Neuroinflammation.

    Science.gov (United States)

    Watson, P Marc D; Kavanagh, Edel; Allenby, Gary; Vassey, Matthew

    2017-02-01

    Neurodegeneration and neuroinflammation are key features in a range of chronic central nervous system (CNS) diseases such as Alzheimer's and Parkinson's disease, as well as acute conditions like stroke and traumatic brain injury, for which there remains significant unmet clinical need. It is now well recognized that current cell culture methodologies are limited in their ability to recapitulate the cellular environment that is present in vivo, and there is a growing body of evidence to show that three-dimensional (3D) culture systems represent a more physiologically accurate model than traditional two-dimensional (2D) cultures. Given the complexity of the environment from which cells originate, and their various cell-cell and cell-matrix interactions, it is important to develop models that can be controlled and reproducible for drug discovery. 3D cell models have now been developed for almost all CNS cell types, including neurons, astrocytes, microglia, and oligodendrocyte cells. This review will highlight a number of current and emerging techniques for the culture of astrocytes and microglia, glial cell types with a critical role in neurodegenerative and neuroinflammatory conditions. We describe recent advances in glial cell culture using electrospun polymers and hydrogel macromolecules, and highlight how these novel culture environments influence astrocyte and microglial phenotypes in vitro, as compared to traditional 2D systems. These models will be explored to illuminate current trends in the techniques used to create 3D environments for application in research and drug discovery focused on astrocytes and microglial cells.

  2. Optimization of liquid overlay technique to formulate heterogenic 3D co-cultures models.

    Science.gov (United States)

    Costa, Elisabete C; Gaspar, Vítor M; Coutinho, Paula; Correia, Ilídio J

    2014-08-01

    Three-dimensional (3D) cell culture models of solid tumors are currently having a tremendous impact in the in vitro screening of candidate anti-tumoral therapies. These 3D models provide more reliable results than those provided by standard 2D in vitro cell cultures. However, 3D manufacturing techniques need to be further optimized in order to increase the robustness of these models and provide data that can be properly correlated with the in vivo situation. Therefore, in the present study the parameters used for producing multicellular tumor spheroids (MCTS) by liquid overlay technique (LOT) were optimized in order to produce heterogeneous cellular agglomerates comprised of cancer cells and stromal cells, during long periods. Spheroids were produced under highly controlled conditions, namely: (i) agarose coatings; (ii) horizontal stirring, and (iii) a known initial cell number. The simultaneous optimization of these parameters promoted the assembly of 3D characteristic cellular organization similar to that found in the in vivo solid tumors. Such improvements in the LOT technique promoted the assembly of highly reproducible, individual 3D spheroids, with a low cost of production and that can be used for future in vitro drug screening assays.

  3. 3D hepatic cultures simultaneously maintain primary hepatocyte and liver sinusoidal endothelial cell phenotypes.

    Directory of Open Access Journals (Sweden)

    Yeonhee Kim

    Full Text Available Developing in vitro engineered hepatic tissues that exhibit stable phenotype is a major challenge in the field of hepatic tissue engineering. However, the rapid dedifferentiation of hepatic parenchymal (hepatocytes and non-parenchymal (liver sinusoidal endothelial, LSEC cell types when removed from their natural environment in vivo remains a major obstacle. The primary goal of this study was to demonstrate that hepatic cells cultured in layered architectures could preserve or potentially enhance liver-specific behavior of both cell types. Primary rat hepatocytes and rat LSECs (rLSECs were cultured in a layered three-dimensional (3D configuration. The cell layers were separated by a chitosan-hyaluronic acid polyelectrolyte multilayer (PEM, which served to mimic the Space of Disse. Hepatocytes and rLSECs exhibited several key phenotypic characteristics over a twelve day culture period. Immunostaining for the sinusoidal endothelial 1 antibody (SE-1 demonstrated that rLSECs cultured in the 3D hepatic model maintained this unique feature over twelve days. In contrast, rLSECs cultured in monolayers lost their phenotype within three days. The unique stratified structure of the 3D culture resulted in enhanced heterotypic cell-cell interactions, which led to improvements in hepatocyte functions. Albumin production increased three to six fold in the rLSEC-PEM-Hepatocyte cultures. Only rLSEC-PEM-Hepatocyte cultures exhibited increasing CYP1A1/2 and CYP3A activity. Well-defined bile canaliculi were observed only in the rLSEC-PEM-Hepatocyte cultures. Together, these data suggest that rLSEC-PEM-Hepatocyte cultures are highly suitable models to monitor the transformation of toxins in the liver and their transport out of this organ. In summary, these results indicate that the layered rLSEC-PEM-hepatocyte model, which recapitulates key features of hepatic sinusoids, is a potentially powerful medium for obtaining comprehensive knowledge on liver metabolism

  4. Integrated Data Capturing Requirements for 3d Semantic Modelling of Cultural Heritage: the Inception Protocol

    Science.gov (United States)

    Di Giulio, R.; Maietti, F.; Piaia, E.; Medici, M.; Ferrari, F.; Turillazzi, B.

    2017-02-01

    The generation of high quality 3D models can be still very time-consuming and expensive, and the outcome of digital reconstructions is frequently provided in formats that are not interoperable, and therefore cannot be easily accessed. This challenge is even more crucial for complex architectures and large heritage sites, which involve a large amount of data to be acquired, managed and enriched by metadata. In this framework, the ongoing EU funded project INCEPTION - Inclusive Cultural Heritage in Europe through 3D semantic modelling proposes a workflow aimed at the achievements of efficient 3D digitization methods, post-processing tools for an enriched semantic modelling, web-based solutions and applications to ensure a wide access to experts and non-experts. In order to face these challenges and to start solving the issue of the large amount of captured data and time-consuming processes in the production of 3D digital models, an Optimized Data Acquisition Protocol (DAP) has been set up. The purpose is to guide the processes of digitization of cultural heritage, respecting needs, requirements and specificities of cultural assets.

  5. A 3D human neural cell culture system for modeling Alzheimer’s disease

    Science.gov (United States)

    Kim, Young Hye; Choi, Se Hoon; D’Avanzo, Carla; Hebisch, Matthias; Sliwinski, Christopher; Bylykbashi, Enjana; Washicosky, Kevin J.; Klee, Justin B.; Brüstle, Oliver; Tanzi, Rudolph E.; Kim, Doo Yeon

    2015-01-01

    Stem cell technologies have facilitated the development of human cellular disease models that can be used to study pathogenesis and test therapeutic candidates. These models hold promise for complex neurological diseases such as Alzheimer’s disease (AD) because existing animal models have been unable to fully recapitulate all aspects of pathology. We recently reported the characterization of a novel three-dimensional (3D) culture system that exhibits key events in AD pathogenesis, including extracellular aggregation of β-amyloid and accumulation of hyperphosphorylated tau. Here we provide instructions for the generation and analysis of 3D human neural cell cultures, including the production of genetically modified human neural progenitor cells (hNPCs) with familial AD mutations, the differentiation of the hNPCs in a 3D matrix, and the analysis of AD pathogenesis. The 3D culture generation takes 1–2 days. The aggregation of β-amyloid is observed after 6-weeks of differentiation followed by robust tau pathology after 10–14 weeks. PMID:26068894

  6. Comparative analysis of 3D culture methods on human HepG2 cells.

    Science.gov (United States)

    Luckert, Claudia; Schulz, Christina; Lehmann, Nadja; Thomas, Maria; Hofmann, Ute; Hammad, Seddik; Hengstler, Jan G; Braeuning, Albert; Lampen, Alfonso; Hessel, Stefanie

    2017-01-01

    Human primary hepatocytes represent a gold standard in in vitro liver research. Due to their low availability and high costs alternative liver cell models with comparable morphological and biochemical characteristics have come into focus. The human hepatocarcinoma cell line HepG2 is often used as a liver model for toxicity studies. However, under two-dimensional (2D) cultivation conditions the expression of xenobiotic-metabolizing enzymes and typical liver markers such as albumin is very low. Cultivation for 21 days in a three-dimensional (3D) Matrigel culture system has been reported to strongly increase the metabolic competence of HepG2 cells. In our present study we further compared HepG2 cell cultivation in three different 3D systems: collagen, Matrigel and Alvetex culture. Cell morphology, albumin secretion, cytochrome P450 monooxygenase enzyme activities, as well as gene expression of xenobiotic-metabolizing and liver-specific enzymes were analyzed after 3, 7, 14, and 21 days of cultivation. Our results show that the previously reported increase of metabolic competence of HepG2 cells is not primarily the result of 3D culture but a consequence of the duration of cultivation. HepG2 cells grown for 21 days in 2D monolayer exhibit comparable biochemical characteristics, CYP activities and gene expression patterns as all 3D culture systems used in our study. However, CYP activities did not reach the level of HepaRG cells. In conclusion, the increase of metabolic competence of the hepatocarcinoma cell line HepG2 is not due to 3D cultivation but rather a result of prolonged cultivation time.

  7. An Air-Liquid Interface Culture System for 3D Organoid Culture of Diverse Primary Gastrointestinal Tissues.

    Science.gov (United States)

    Li, Xingnan; Ootani, Akifumi; Kuo, Calvin

    2016-01-01

    Conventional in vitro analysis of gastrointestinal epithelium usually relies on two-dimensional (2D) culture of epithelial cell lines as monolayer on impermeable surfaces. However, the lack of context of differentiation and tissue architecture in 2D culture can hinder the faithful recapitulation of the phenotypic and morphological characteristics of native epithelium. Here, we describe a robust long-term three-dimensional (3D) culture methodology for gastrointestinal culture, which incorporates both epithelial and mesenchymal/stromal components into a collagen-based air-liquid interface 3D culture system. This system allows vigorously expansion of primary gastrointestinal epithelium for over 60 days as organoids with both proliferation and multilineage differentiation, indicating successful long-term intestinal culture within a microenvironment accurately recapitulating the stem cell niche.

  8. Registration of 3D and Multispectral Data for the Study of Cultural Heritage Surfaces

    Directory of Open Access Journals (Sweden)

    Frank Boochs

    2013-01-01

    Full Text Available We present a technique for the multi-sensor registration of featureless datasets based on the photogrammetric tracking of the acquisition systems in use. This method is developed for the in situ study of cultural heritage objects and is tested by digitizing a small canvas successively with a 3D digitization system and a multispectral camera while simultaneously tracking the acquisition systems with four cameras and using a cubic target frame with a side length of 500 mm. The achieved tracking accuracy is better than 0.03 mm spatially and 0.150 mrad angularly. This allows us to seamlessly register the 3D acquisitions and to project the multispectral acquisitions on the 3D model.

  9. Spheroid culture as a tool for creating 3D complex tissues.

    Science.gov (United States)

    Fennema, Eelco; Rivron, Nicolas; Rouwkema, Jeroen; van Blitterswijk, Clemens; de Boer, Jan

    2013-02-01

    3D cell culture methods confer a high degree of clinical and biological relevance to in vitro models. This is specifically the case with the spheroid culture, where a small aggregate of cells grows free of foreign materials. In spheroid cultures, cells secrete the extracellular matrix (ECM) in which they reside, and they can interact with cells from their original microenvironment. The value of spheroid cultures is increasing quickly due to novel microfabricated platforms amenable to high-throughput screening (HTS) and advances in cell culture. Here, we review new possibilities that combine the strengths of spheroid culture with new microenvironment fabrication methods that allow for the creation of large numbers of highly reproducible, complex tissues.

  10. 3D MODELLING AND INTERACTIVE WEB-BASED VISUALIZATION OF CULTURAL HERITAGE OBJECTS

    Directory of Open Access Journals (Sweden)

    M. N. Koeva

    2016-06-01

    Full Text Available Nowadays, there are rapid developments in the fields of photogrammetry, laser scanning, computer vision and robotics, together aiming to provide highly accurate 3D data that is useful for various applications. In recent years, various LiDAR and image-based techniques have been investigated for 3D modelling because of their opportunities for fast and accurate model generation. For cultural heritage preservation and the representation of objects that are important for tourism and their interactive visualization, 3D models are highly effective and intuitive for present-day users who have stringent requirements and high expectations. Depending on the complexity of the objects for the specific case, various technological methods can be applied. The selected objects in this particular research are located in Bulgaria – a country with thousands of years of history and cultural heritage dating back to ancient civilizations. \\this motivates the preservation, visualisation and recreation of undoubtedly valuable historical and architectural objects and places, which has always been a serious challenge for specialists in the field of cultural heritage. In the present research, comparative analyses regarding principles and technological processes needed for 3D modelling and visualization are presented. The recent problems, efforts and developments in interactive representation of precious objects and places in Bulgaria are presented. Three technologies based on real projects are described: (1 image-based modelling using a non-metric hand-held camera; (2 3D visualization based on spherical panoramic images; (3 and 3D geometric and photorealistic modelling based on architectural CAD drawings. Their suitability for web-based visualization are demonstrated and compared. Moreover the possibilities for integration with additional information such as interactive maps, satellite imagery, sound, video and specific information for the objects are described. This

  11. Deploying 3D technologies for the documentation of tangible cultural heritage

    Directory of Open Access Journals (Sweden)

    Karina Rodríguez Echavarria

    2012-05-01

    Full Text Available Cada vez más, los visitantes y profesionistas del patrimonio cultural esperan más de las tecnologías 3D. Es por esto que el proyecto 3D-COFORM pretende hacer que la tecnología 3D sea una realidad para el sector. El proyecto está desarrollando herramientas dirigidas a diversos tipos de usuarios y a la vez investigando sobre las cuestiones prácticas para su implementación en organizaciones de patrimonio. En esta comunicación se describe la metodología para lograr esto, así como diferentes tipos de pruebas llevadas a cabo por el proyecto. También propone tres modelos para la implementación de las tecnologías 3D y describe una de ellas con más detalle. Aunque estos resultados son preliminares, se espera que contribuyan a que el sector vea la implementación de tecnologías 3D como una opción sustentable.

  12. Mackay campus of environmental education and digital cultural construction: the application of 3D virtual reality

    Science.gov (United States)

    Chien, Shao-Chi; Chung, Yu-Wei; Lin, Yi-Hsuan; Huang, Jun-Yi; Chang, Jhih-Ting; He, Cai-Ying; Cheng, Yi-Wen

    2012-04-01

    This study uses 3D virtual reality technology to create the "Mackay campus of the environmental education and digital cultural 3D navigation system" for local historical sites in the Tamsui (Hoba) area, in hopes of providing tourism information and navigation through historical sites using a 3D navigation system. We used Auto CAD, Sketch Up, and SpaceEyes 3D software to construct the virtual reality scenes and create the school's historical sites, such as the House of Reverends, the House of Maidens, the Residence of Mackay, and the Education Hall. We used this technology to complete the environmental education and digital cultural Mackay campus . The platform we established can indeed achieve the desired function of providing tourism information and historical site navigation. The interactive multimedia style and the presentation of the information will allow users to obtain a direct information response. In addition to showing the external appearances of buildings, the navigation platform can also allow users to enter the buildings to view lifelike scenes and textual information related to the historical sites. The historical sites are designed according to their actual size, which gives users a more realistic feel. In terms of the navigation route, the navigation system does not force users along a fixed route, but instead allows users to freely control the route they would like to take to view the historical sites on the platform.

  13. Conducting Polymer Scaffolds for Hosting and Monitoring 3D Cell Culture

    KAUST Repository

    Inal, Sahika

    2017-05-03

    This work reports the design of a live-cell monitoring platform based on a macroporous scaffold of a conducting polymer, poly(3,4-ethylene dioxythiophene):poly(styrenesulfonate). The conducting polymer scaffolds support 3D cell cultures due to their biocompatibility and tissue-like elasticity, which can be manipulated by inclusion of biopolymers such as collagen. Integration of a media perfusion tube inside the scaffold enables homogenous cell spreading and fluid transport throughout the scaffold, ensuring long term cell viability. This also allows for co-culture of multiple cell types inside the scaffold. The inclusion of cells within the porous architecture affects the impedance of the electrically conducting polymer network and, thus, is utilized as an in situ tool to monitor cell growth. Therefore, while being an integral part of the 3D tissue, the conducting polymer is an active component, enhancing the tissue function, and forming the basis for a bioelectronic device with integrated sensing capability.

  14. 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.

  15. Differentiation capacity and maintenance of differentiated phenotypes of human mesenchymal stromal cells cultured on two distinct types of 3D polymeric scaffolds.

    Science.gov (United States)

    Leferink, A M; Santos, D; Karperien, M; Truckenmüller, R K; van Blitterswijk, C A; Moroni, L

    2015-12-01

    Many studies have shown the influence of soluble factors and material properties on the differentiation capacity of mesenchymal stromal cells (MSCs) cultured as monolayers. These types of two-dimensional (2D) studies can be used as simplified models to understand cell processes related to stem cell sensing and mechano-transduction in a three-dimensional (3D) context. For several other mechanisms such as cell-cell signaling, cell proliferation and cell morphology, it is well-known that cells behave differently on a planar surface compared to cells in 3D environments. In classical tissue engineering approaches, a combination of cells, 3D scaffolds and soluble factors are considered as the key ingredients for the generation of mechanically stable 3D tissue constructs. However, when MSCs are used for tissue engineering strategies, little is known about the maintenance of their differentiation potential in 3D scaffolds after the removal of differentiation soluble factors. In this study, the differentiation potential of human MSCs (hMSCs) into the chondrogenic and osteogenic lineages on two distinct 3D scaffolds, additive manufactured electrospun scaffolds, was assessed and compared to conventional 2D culture. Human MSCs cultured in the presence of soluble factors in 3D showed to differentiate to the same extent as hMSCs cultured as 2D monolayers or as scaffold-free pellets, indicating that the two scaffolds do not play a consistent role in the differentiation process. In the case of phenotypic changes, the achieved differentiated phenotype was not maintained after the removal of soluble factors, suggesting that the plasticity of hMSCs is retained in 3D cell culture systems. This finding can have implications for future tissue engineering approaches in which the validation of hMSC differentiation on 3D scaffolds will not be sufficient to ensure the maintenance of the functionality of the cells in the absence of appropriate differentiation signals.

  16. Culturing and applications of rotating wall vessel bioreactor derived 3D epithelial cell models.

    Science.gov (United States)

    Radtke, Andrea L; Herbst-Kralovetz, Melissa M

    2012-04-03

    Cells and tissues in the body experience environmental conditions that influence their architecture, intercellular communications, and overall functions. For in vitro cell culture models to accurately mimic the tissue of interest, the growth environment of the culture is a critical aspect to consider. Commonly used conventional cell culture systems propagate epithelial cells on flat two-dimensional (2-D) impermeable surfaces. Although much has been learned from conventional cell culture systems, many findings are not reproducible in human clinical trials or tissue explants, potentially as a result of the lack of a physiologically relevant microenvironment. Here, we describe a culture system that overcomes many of the culture condition boundaries of 2-D cell cultures, by using the innovative rotating wall vessel (RWV) bioreactor technology. We and others have shown that organotypic RWV-derived models can recapitulate structure, function, and authentic human responses to external stimuli similarly to human explant tissues (1-6). The RWV bioreactor is a suspension culture system that allows for the growth of epithelial cells under low physiological fluid shear conditions. The bioreactors come in two different formats, a high-aspect rotating vessel (HARV) or a slow-turning lateral vessel (STLV), in which they differ by their aeration source. Epithelial cells are added to the bioreactor of choice in combination with porous, collagen-coated microcarrier beads (Figure 1A). The cells utilize the beads as a growth scaffold during the constant free fall in the bioreactor (Figure 1B). The microenvironment provided by the bioreactor allows the cells to form three-dimensional (3-D) aggregates displaying in vivo-like characteristics often not observed under standard 2-D culture conditions (Figure 1D). These characteristics include tight junctions, mucus production, apical/basal orientation, in vivo protein localization, and additional epithelial cell-type specific properties

  17. Universal lab-on-a-chip platform for complex, perfused 3D cell cultures

    Science.gov (United States)

    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.

  18. The Rise of Avatars in Positive Psychology: Enabling Military Cultures in a 3D World for Strengths of Character Development

    Science.gov (United States)

    2010-03-01

    Rethinking Transfer:  A  Simple Proposal With Multiple Implications,” Review of  Research in  Education , 24(61), (1999): 83.  v  Jerome   Bruner , The Culture...Policy, National Defense  University.  Huntington,  S . P.  (1985).  The soldier and the  state:  The  theory  and practice of civil‐ military relations...Cultures in a 3D World for Strengths of Character Development 5a. CONTRACT NUMBER 5b. GRANT NUMBER 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR( S ) 5d. PROJECT

  19. Segmentation of 3d Models for Cultural Heritage Structural Analysis - Some Critical Issues

    Science.gov (United States)

    Gonizzi Barsanti, S.; Guidi, G.; De Luca, L.

    2017-08-01

    Cultural Heritage documentation and preservation has become a fundamental concern in this historical period. 3D modelling offers a perfect aid to record ancient buildings and artefacts and can be used as a valid starting point for restoration, conservation and structural analysis, which can be performed by using Finite Element Methods (FEA). The models derived from reality-based techniques, made up of the exterior surfaces of the objects captured at high resolution, are - for this reason - made of millions of polygons. Such meshes are not directly usable in structural analysis packages and need to be properly pre-processed in order to be transformed in volumetric meshes suitable for FEA. In addition, dealing with ancient objects, a proper segmentation of 3D volumetric models is needed to analyse the behaviour of the structure with the most suitable level of detail for the different sections of the structure under analysis. Segmentation of 3D models is still an open issue, especially when dealing with ancient, complicated and geometrically complex objects that imply the presence of anomalies and gaps, due to environmental agents such as earthquakes, pollution, wind and rain, or human factors. The aims of this paper is to critically analyse some of the different methodologies and algorithms available to segment a 3D point cloud or a mesh, identifying difficulties and problems by showing examples on different structures.

  20. Assembly of Hepatocyte Spheroids Using Magnetic 3D Cell Culture for CYP450 Inhibition/Induction

    Directory of Open Access Journals (Sweden)

    Pujan K. Desai

    2017-05-01

    Full Text Available There is a significant need for in vitro methods to study drug-induced liver injury that are rapid, reproducible, and scalable for existing high-throughput systems. However, traditional monolayer and suspension cultures of hepatocytes are difficult to handle and risk the loss of phenotype. Generally, three-dimensional (3D cell culture platforms help recapitulate native liver tissue phenotype, but suffer from technical limitations for high-throughput screening, including scalability, speed, and handling. Here, we developed a novel assay for cytochrome P450 (CYP450 induction/inhibition using magnetic 3D cell culture that overcomes the limitations of other platforms by aggregating magnetized cells with magnetic forces. With this platform, spheroids can be rapidly assembled and easily handled, while replicating native liver function. We assembled spheroids of primary human hepatocytes in a 384-well format and maintained this culture over five days, including a 72 h induction period with known CYP450 inducers/inhibitors. CYP450 activity and viability in the spheroids were assessed and compared in parallel with monolayers. CYP450 activity was induced/inhibited in spheroids as expected, separate from any toxic response. Spheroids showed a significantly higher baseline level of CYP450 activity and induction over monolayers. Positive staining in spheroids for albumin and multidrug resistance-associated protein (MRP2 indicates the preservation of hepatocyte function within spheroids. The study presents a proof-of-concept for the use of magnetic 3D cell culture for the assembly and handling of novel hepatic tissue models.

  1. 3D correlative light and electron microscopy of cultured cells using serial blockface scanning electron microscopy

    Science.gov (United States)

    Lerner, Thomas R.; Burden, Jemima J.; Nkwe, David O.; Pelchen-Matthews, Annegret; Domart, Marie-Charlotte; Durgan, Joanne; Weston, Anne; Jones, Martin L.; Peddie, Christopher J.; Carzaniga, Raffaella; Florey, Oliver; Marsh, Mark; Gutierrez, Maximiliano G.

    2017-01-01

    ABSTRACT The processes of life take place in multiple dimensions, but imaging these processes in even three dimensions is challenging. Here, we describe a workflow for 3D correlative light and electron microscopy (CLEM) of cell monolayers using fluorescence microscopy to identify and follow biological events, combined with serial blockface scanning electron microscopy to analyse the underlying ultrastructure. The workflow encompasses all steps from cell culture to sample processing, imaging strategy, and 3D image processing and analysis. We demonstrate successful application of the workflow to three studies, each aiming to better understand complex and dynamic biological processes, including bacterial and viral infections of cultured cells and formation of entotic cell-in-cell structures commonly observed in tumours. Our workflow revealed new insight into the replicative niche of Mycobacterium tuberculosis in primary human lymphatic endothelial cells, HIV-1 in human monocyte-derived macrophages, and the composition of the entotic vacuole. The broad application of this 3D CLEM technique will make it a useful addition to the correlative imaging toolbox for biomedical research. PMID:27445312

  2. 3D correlative light and electron microscopy of cultured cells using serial blockface scanning electron microscopy.

    Science.gov (United States)

    Russell, Matthew R G; Lerner, Thomas R; Burden, Jemima J; Nkwe, David O; Pelchen-Matthews, Annegret; Domart, Marie-Charlotte; Durgan, Joanne; Weston, Anne; Jones, Martin L; Peddie, Christopher J; Carzaniga, Raffaella; Florey, Oliver; Marsh, Mark; Gutierrez, Maximiliano G; Collinson, Lucy M

    2017-01-01

    The processes of life take place in multiple dimensions, but imaging these processes in even three dimensions is challenging. Here, we describe a workflow for 3D correlative light and electron microscopy (CLEM) of cell monolayers using fluorescence microscopy to identify and follow biological events, combined with serial blockface scanning electron microscopy to analyse the underlying ultrastructure. The workflow encompasses all steps from cell culture to sample processing, imaging strategy, and 3D image processing and analysis. We demonstrate successful application of the workflow to three studies, each aiming to better understand complex and dynamic biological processes, including bacterial and viral infections of cultured cells and formation of entotic cell-in-cell structures commonly observed in tumours. Our workflow revealed new insight into the replicative niche of Mycobacterium tuberculosis in primary human lymphatic endothelial cells, HIV-1 in human monocyte-derived macrophages, and the composition of the entotic vacuole. The broad application of this 3D CLEM technique will make it a useful addition to the correlative imaging toolbox for biomedical research. © 2017. Published by The Company of Biologists Ltd.

  3. Video lensfree microscopy of 2D and 3D culture of cells

    Science.gov (United States)

    Allier, C. P.; Vinjimore Kesavan, S.; Coutard, J.-G.; Cioni, O.; Momey, F.; Navarro, F.; Menneteau, M.; Chalmond, B.; Obeid, P.; Haguet, V.; David-Watine, B.; Dubrulle, N.; Shorte, S.; van der Sanden, B.; Di Natale, C.; Hamard, L.; Wion, D.; Dolega, M. E.; Picollet-D'hahan, N.; Gidrol, X.; Dinten, J.-M.

    2014-03-01

    Innovative imaging methods are continuously developed to investigate the function of biological systems at the microscopic scale. As an alternative to advanced cell microscopy techniques, we are developing lensfree video microscopy that opens new ranges of capabilities, in particular at the mesoscopic level. Lensfree video microscopy allows the observation of a cell culture in an incubator over a very large field of view (24 mm2) for extended periods of time. As a result, a large set of comprehensive data can be gathered with strong statistics, both in space and time. Video lensfree microscopy can capture images of cells cultured in various physical environments. We emphasize on two different case studies: the quantitative analysis of the spontaneous network formation of HUVEC endothelial cells, and by coupling lensfree microscopy with 3D cell culture in the study of epithelial tissue morphogenesis. In summary, we demonstrate that lensfree video microscopy is a powerful tool to conduct cell assays in 2D and 3D culture experiments. The applications are in the realms of fundamental biology, tissue regeneration, drug development and toxicology studies.

  4. 2D and 3D Self-Assembling Nanofiber Hydrogels for Cardiomyocyte Culture

    Directory of Open Access Journals (Sweden)

    Liisa Ikonen

    2013-01-01

    Full Text Available Collagen is a widely used biomaterial in cardiac tissue engineering studies. However, as a natural material, it suffers from variability between batches that can complicate the standardization of culture conditions. In contrast, synthetic materials are modifiable, have well-defined structures and more homogeneous batches can be produced. In this study, several collagen-like synthetic self-assembling nanofiber hydrogels were examined for their suitability for cardiomyocyte culture in 2D and 3D. Six different nanofiber coatings were used in the 2D format with neonatal rat cardiomyocytes (NRCs and human embryonic stem-cell-derived cardiomyocytes (hESC-CMs. The viability, growth, and functionality of the 2D-cultured cardiomyocytes were evaluated. The best-performing nanofiber coatings were selected for 3D experiments. Hydrophilic pH-sensitive nanofiber hydrogel coassembled with hyaluronic acid performed best with both NRCs and hESC-CMs. Hydrophilic non-pH-sensitive nanofiber hydrogels supported the growth of NRCs; however, their ability to promote attachment and growth of hESC-CMs was limited. NRCs also grew on hydrophobic nanofiber hydrogels; however, the cell-supporting capacity of these hydrogels was inferior to that of the hydrophilic hydrogel materials. This is the first study demonstrating that hydrophilic self-assembling nanofiber hydrogels support the culture of both NRCs and hESC-CMs, which suggests that these biomaterials hold promise for cardiac tissue engineering.

  5. Fibroblasts Influence Survival and Therapeutic Response in a 3D Co-Culture Model.

    Science.gov (United States)

    Majety, Meher; Pradel, Leon P; Gies, Manuela; Ries, Carola H

    2015-01-01

    In recent years, evidence has indicated that the tumor microenvironment (TME) plays a significant role in tumor progression. Fibroblasts represent an abundant cell population in the TME and produce several growth factors and cytokines. Fibroblasts generate a suitable niche for tumor cell survival and metastasis under the influence of interactions between fibroblasts and tumor cells. Investigating these interactions requires suitable experimental systems to understand the cross-talk involved. Most in vitro experimental systems use 2D cell culture and trans-well assays to study these interactions even though these paradigms poorly represent the tumor, in which direct cell-cell contacts in 3D spaces naturally occur. Investigating these interactions in vivo is of limited value due to problems regarding the challenges caused by the species-specificity of many molecules. Thus, it is essential to use in vitro models in which human fibroblasts are co-cultured with tumor cells to understand their interactions. Here, we developed a 3D co-culture model that enables direct cell-cell contacts between pancreatic, breast and or lung tumor cells and human fibroblasts/ or tumor-associated fibroblasts (TAFs). We found that co-culturing with fibroblasts/TAFs increases the proliferation in of several types of cancer cells. We also observed that co-culture induces differential expression of soluble factors in a cancer type-specific manner. Treatment with blocking antibodies against selected factors or their receptors resulted in the inhibition of cancer cell proliferation in the co-cultures. Using our co-culture model, we further revealed that TAFs can influence the response to therapeutic agents in vitro. We suggest that this model can be reliably used as a tool to investigate the interactions between a tumor and the TME.

  6. In vitro evaluation of curcumin effects on breast adenocarcinoma 2D and 3D cell cultures.

    Science.gov (United States)

    Abuelba, Hussam; Cotrutz, Carmen Elena; Stoica, Bogdan Alexandru; Stoica, Laura; Olinici, DoiniŢa; Petreuş, Tudor

    2015-01-01

    Breast adenocarcinoma cell line MDA-MB-231, even if it expresses low levels of E-cadherin, still readily form multicellular aggregates of cells, namely spheroids. Curcumin is a diarylheptanoid antitumoral drug while it significantly inhibits cell migration, invasion, and colony formation in vitro and reduces tumor growth and liver metastasis in vivo. Curcumin photoactivation may enhance antiapoptotic role against cancer cells. To evaluate the effect of low curcumin concentrations, ranged from 1.9 to 15 μM, with and without photoactivation, using a manufactured 670 nm LED-matrix. A secondary aim was to evaluate the ideal method to produce easy-to-use tumor cell spheroids, comparing two low adherence plate supports. Breast adenocarcinoma cell line MDA-MB-231 were cultured according to 2D monolayer and 3D spheroid models then submitted to normal and photoactivated curcumin in micromolar concentrations. MTT assay was used to evaluate cell viability following curcumin application on cells. On 2D cell cultures, curcumin inhibits cell tumor development and proliferation at concentrations of 15 μM, with a viability of 65.7% at 48 hours incubation time. A decreased viability up to 25% for a concentration of 15 μM was recorded following photoactivation and cytotoxic action on breast cancer tumor cell line continued at concentrations of 7.5 and 3.75 μM. Curcumin photoactivation increases pro-apoptotic effects in both 2D and 3D tumor cell culture models and also responsiveness to curcumin is slightly reduced in spheroid-like structures. Thus, 3D tumor cell culture systems appear to be the ideal environment for in vitro assays regarding anticancer drug effects on cell viability.

  7. Minimal camera networks for 3D image based modeling of cultural heritage objects.

    Science.gov (United States)

    Alsadik, Bashar; Gerke, Markus; Vosselman, George; Daham, Afrah; Jasim, Luma

    2014-03-25

    3D modeling of cultural heritage objects like artifacts, statues and buildings is nowadays an important tool for virtual museums, preservation and restoration. In this paper, we introduce a method to automatically design a minimal imaging network for the 3D modeling of cultural heritage objects. This becomes important for reducing the image capture time and processing when documenting large and complex sites. Moreover, such a minimal camera network design is desirable for imaging non-digitally documented artifacts in museums and other archeological sites to avoid disturbing the visitors for a long time and/or moving delicate precious objects to complete the documentation task. The developed method is tested on the Iraqi famous statue "Lamassu". Lamassu is a human-headed winged bull of over 4.25 m in height from the era of Ashurnasirpal II (883-859 BC). Close-range photogrammetry is used for the 3D modeling task where a dense ordered imaging network of 45 high resolution images were captured around Lamassu with an object sample distance of 1 mm. These images constitute a dense network and the aim of our study was to apply our method to reduce the number of images for the 3D modeling and at the same time preserve pre-defined point accuracy. Temporary control points were fixed evenly on the body of Lamassu and measured by using a total station for the external validation and scaling purpose. Two network filtering methods are implemented and three different software packages are used to investigate the efficiency of the image orientation and modeling of the statue in the filtered (reduced) image networks. Internal and external validation results prove that minimal image networks can provide highly accurate records and efficiency in terms of visualization, completeness, processing time (>60% reduction) and the final accuracy of 1 mm.

  8. Minimal Camera Networks for 3D Image Based Modeling of Cultural Heritage Objects

    Directory of Open Access Journals (Sweden)

    Bashar Alsadik

    2014-03-01

    Full Text Available 3D modeling of cultural heritage objects like artifacts, statues and buildings is nowadays an important tool for virtual museums, preservation and restoration. In this paper, we introduce a method to automatically design a minimal imaging network for the 3D modeling of cultural heritage objects. This becomes important for reducing the image capture time and processing when documenting large and complex sites. Moreover, such a minimal camera network design is desirable for imaging non-digitally documented artifacts in museums and other archeological sites to avoid disturbing the visitors for a long time and/or moving delicate precious objects to complete the documentation task. The developed method is tested on the Iraqi famous statue “Lamassu”. Lamassu is a human-headed winged bull of over 4.25 m in height from the era of Ashurnasirpal II (883–859 BC. Close-range photogrammetry is used for the 3D modeling task where a dense ordered imaging network of 45 high resolution images were captured around Lamassu with an object sample distance of 1 mm. These images constitute a dense network and the aim of our study was to apply our method to reduce the number of images for the 3D modeling and at the same time preserve pre-defined point accuracy. Temporary control points were fixed evenly on the body of Lamassu and measured by using a total station for the external validation and scaling purpose. Two network filtering methods are implemented and three different software packages are used to investigate the efficiency of the image orientation and modeling of the statue in the filtered (reduced image networks. Internal and external validation results prove that minimal image networks can provide highly accurate records and efficiency in terms of visualization, completeness, processing time (>60% reduction and the final accuracy of 1 mm.

  9. A novel MCF-10A line allowing conditional oncogene expression in 3D culture

    Directory of Open Access Journals (Sweden)

    Danke Christina

    2011-07-01

    Full Text Available Introduction Non-transformed mammary epithelial cell lines such as MCF-10A recapitulate epithelial morphogenesis in three-dimensional (3D tissue culture by forming acinar structures. They represent an important tool to characterize the biological properties of oncogenes and to model early carcinogenic events. So far, however, these approaches were restricted to cells with constitutive oncogene expression prior to the set-up of 3D cultures. Although very informative, this experimental setting has precluded the analysis of effects caused by sudden oncoprotein expression or withdrawal in established epithelial cultures. Here, we report the establishment and use of a stable MCF-10A cell line (MCF-10Atet fitted with a novel and improved doxycycline (dox-regulated expression system allowing the conditional expression of any transgene. Methods MCF-10Atet cells were generated by stable transfection with pWHE644, a vector expressing a second generation tetracycline-regulated transactivator and a novel transcriptional silencer. In order to test the properties of this new repressor/activator switch, MCF-10Atet cells were transfected with a second plasmid, pTET-HABRAF-IRES-GFP, which responds to dox treatment with the production of a bi-cistronic transcript encoding hemagglutinin-tagged B-Raf and green fluorescent protein (GFP. This improved conditional expression system was then characterized in detail in terms of its response to various dox concentrations and exposure times. The plasticity of the phenotype provoked by oncogenic B-RafV600E in MCF-10Atet cells was analyzed in 3D cultures by dox exposure and subsequent wash-out. Results MCF-10Atet cells represent a tightly controlled, conditional gene expression system. Using B-RafV600E as a model oncoprotein, we show that its sudden expression in established 3D cultures results in the loss of acinar organization, the induction of an invasive phenotype and hallmarks of epithelial-to-mesenchymal transition

  10. Engineering Cellular Microenvironments with Photo- and Enzymatically Responsive Hydrogels: Toward Biomimetic 3D Cell Culture Models.

    Science.gov (United States)

    Tam, Roger Y; Smith, Laura J; Shoichet, Molly S

    2017-04-18

    Conventional cell culture techniques using 2D polystyrene or glass have provided great insight into key biochemical mechanisms responsible for cellular events such as cell proliferation, differentiation, and cell-cell interactions. However, the physical and chemical properties of 2D culture in vitro are dramatically different than those found in the native cellular microenvironment in vivo. Cells grown on 2D substrates differ significantly from those grown in vivo, and this explains, in part, why many promising drug candidates discovered through in vitro drug screening assays fail when they are translated to in vivo animal or human models. To overcome this obstacle, 3D cell culture using biomimetic hydrogels has emerged as an alternative strategy to recapitulate native cell growth in vitro. Hydrogels, which are water-swollen polymers, can be synthetic or naturally derived. Many methods have been developed to control the physical and chemical properties of the hydrogels to match those found in specific tissues. Compared to 2D culture, cells cultured in 3D gels with the appropriate physicochemical cues can behave more like they naturally do in vivo. While conventional hydrogels involve modifications to the bulk material to mimic the static aspects of the cellular microenvironment, recent progress has focused on using more dynamic hydrogels, the chemical and physical properties of which can be altered with external stimuli to better mimic the dynamics of the native cellular microenvironment found in vivo. In this Account, we describe our progress in designing stimuli-responsive, optically transparent hydrogels that can be used as biomimetic extracellular matrices (ECMs) to study cell differentiation and migration in the context of modeling the nervous system and cancer. Specifically, we developed photosensitive agarose and hyaluronic acid hydrogels that are activated by single or two-photon irradiation for biomolecule immobilization at specific volumes within the 3D

  11. Some Experiences in 3D Laser Scanning for Assisting Restoration and Evaluating Damage in Cultural Heritage

    Science.gov (United States)

    Fuentes, L. M.; Finat, Javier; Fernández-Martin, J. J.; Martínez, J.; SanJose, J. I.

    The recent incorporation of laser devices provides advanced tools for assisting the conservation and restoration of Cultural Heritage. It is necessary to have as complete as possible understanding of the object state before evaluating or defining the reach of the restoration process. Thus, a special effort is devoted to surveying, measuring and generating a high-resolution 3D model prior to restoration planning. This work presents results of several experiments performed on damaged pieces for evaluation purposes in Cultural Heritage. Some software tools are applied for carving-work analysis, conservation-state monitoring, and simulation of weathering processes for evaluating temporal changes. In all cases considered, a high resolution information capture has been performed with a laser scanner, the Minolta 910. Our approach is flexible enough to be adapted to other kinds of pieces or Cultural Heritage artefacts, in order to provide an assessment for intervention planning in conservation and restoration tasks.

  12. Presenting Cultural Heritage Landscapes - from GIS via 3d Models to Interactive Presentation Frameworks

    Science.gov (United States)

    Prechtel, N.; Münster, S.; Kröber, C.; Schubert, C.; Schietzold, S.

    2013-07-01

    Two current projects of the authors try to approach cultural heritage landscapes from both cultural sciences and geography through a combination of customised geo-information (GIS) and visualisation/presentation technology. In excess of a mere academic use, easyto- handle virtual 3D web presentations may contribute to knowledge, esteem, commemoration and preservation. The examples relate to pre-historic Scythian burial sites in the South-Siberian Altay Mountains ("Uch Enmek") as well as to a "virtual memorial" of contemporary history ("GEPAM"), a chapter of Jewish prosecution in the "Third Reich", which historically connects the town of Dresden with the Czech Terezin (Theresienstadt). It is common knowledge that a profound understanding of (pre-)historic artefacts and places may reflect a larger environment as well as an individual geographic setting. Coming from this background, the presented projects try to find technical solutions. They start from GIS models and aim at customised interactive presentations of 3D models. In using the latter a widely-spanned public is invited to a land- or townscape of specific cultural importance. The geographic space is thought to work as a door to a repository of educational exhibits under the umbrella of a web application. Within this concept a landscape/townscape also accounts for the time dimension in different scales (time of construction/operation versus actual state, and in sense of a season and time of the day as a principal modulator of visual perception of space).

  13. 3-D nasal cultures: Systems toxicological assessment of a candidate modified-risk tobacco product.

    Science.gov (United States)

    Iskandar, Anita R; Mathis, Carole; Martin, Florian; Leroy, Patrice; Sewer, Alain; Majeed, Shoaib; Kuehn, Diana; Trivedi, Keyur; Grandolfo, Davide; Cabanski, Maciej; Guedj, Emmanuel; Merg, Celine; Frentzel, Stefan; Ivanov, Nikolai V; Peitsch, Manuel C; Hoeng, Julia

    2017-01-01

    In vitro toxicology approaches have evolved from a focus on molecular changes within a cell to understanding of toxicity-related mechanisms in systems that can mimic the in vivo environment. The recent development of three dimensional (3-D) organotypic nasal epithelial culture models offers a physiologically robust system for studying the effects of exposure through inhalation. Exposure to cigarette smoke (CS) is associated with nasal inflammation; thus, the nasal epithelium is relevant for evaluating the pathophysiological impact of CS exposure. The present study investigated further the application of in vitro human 3-D nasal epithelial culture models for toxicological assessment of inhalation exposure. Aligned with 3Rs strategy, this study aimed to explore the relevance of a human 3-D nasal culture model to assess the toxicological impact of aerosols generated from a candidate modified risk tobacco product (cMRTP), the Tobacco Heating System (THS) 2.2, as compared with smoke generated from reference cigarette 3R4F. A series of experimental repetitions, where multiple concentrations of THS2.2 aerosol and 3R4F smoke were applied, were conducted to obtain reproducible measurements to understand the cellular/molecular changes that occur following exposure. In agreement with "Toxicity Testing in the 21st Century - a Vision and a Strategy", this study implemented a systems toxicology approach and found that for all tested concentrations the impact of 3R4F smoke was substantially greater than that of THS2.2 aerosol in terms of cytotoxicity levels, alterations in tissue morphology, secretion of pro-inflammatory mediators, impaired ciliary function, and increased perturbed transcriptomes and miRNA expression profiles.

  14. A 3D co-culture microtissue model of the human placenta for nanotoxicity assessment

    DEFF Research Database (Denmark)

    Muoth, Carina; Wichser, Adrian; Monopoli, Marco;

    2016-01-01

    and functionality of the placental tissue. The effects of NPs on the human placenta are not well studied or understood, and predictive in vitro placenta models to achieve mechanistic insights on NP-placenta interactions are essentially lacking. Using the scaffold-free hanging drop technology, we developed a well-organized...... and highly reproducible 3D co-culture microtissue (MT) model consisting of a core of placental fibroblasts surrounded by a trophoblast cell layer, which resembles the structure of the in vivo placental tissue. We could show that secretion levels of human chorionic gonadotropin (hCG) were significantly higher...

  15. Biodynamic Doppler imaging of subcellular motion inside 3D living tissue culture and biopsies (Conference Presentation)

    Science.gov (United States)

    Nolte, David D.

    2016-03-01

    Biodynamic imaging is an emerging 3D optical imaging technology that probes up to 1 mm deep inside three-dimensional living tissue using short-coherence dynamic light scattering to measure the intracellular motions of cells inside their natural microenvironments. Biodynamic imaging is label-free and non-invasive. The information content of biodynamic imaging is captured through tissue dynamics spectroscopy that displays the changes in the Doppler signatures from intracellular constituents in response to applied compounds. The affected dynamic intracellular mechanisms include organelle transport, membrane undulations, cytoskeletal restructuring, strain at cellular adhesions, cytokinesis, mitosis, exo- and endo-cytosis among others. The development of 3D high-content assays such as biodynamic profiling can become a critical new tool for assessing efficacy of drugs and the suitability of specific types of tissue growth for drug discovery and development. The use of biodynamic profiling to predict clinical outcome of living biopsies to cancer therapeutics can be developed into a phenotypic companion diagnostic, as well as a new tool for therapy selection in personalized medicine. This invited talk will present an overview of the optical, physical and physiological processes involved in biodynamic imaging. Several different biodynamic imaging modalities include motility contrast imaging (MCI), tissue-dynamics spectroscopy (TDS) and tissue-dynamics imaging (TDI). A wide range of potential applications will be described that include process monitoring for 3D tissue culture, drug discovery and development, cancer therapy selection, embryo assessment for in-vitro fertilization and artificial reproductive technologies, among others.

  16. A SEMI-AUTOMATED POINT CLOUD PROCESSING METHODOLOGY FOR 3D CULTURAL HERITAGE DOCUMENTATION

    Directory of Open Access Journals (Sweden)

    C. Ö. Kıvılcım

    2016-06-01

    Full Text Available The preliminary phase in any architectural heritage project is to obtain metric measurements and documentation of the building and its individual elements. On the other hand, conventional measurement techniques require tremendous resources and lengthy project completion times for architectural surveys and 3D model production. Over the past two decades, the widespread use of laser scanning and digital photogrammetry have significantly altered the heritage documentation process. Furthermore, advances in these technologies have enabled robust data collection and reduced user workload for generating various levels of products, from single buildings to expansive cityscapes. More recently, the use of procedural modelling methods and BIM relevant applications for historic building documentation purposes has become an active area of research, however fully automated systems in cultural heritage documentation still remains open. In this paper, we present a semi-automated methodology, for 3D façade modelling of cultural heritage assets based on parametric and procedural modelling techniques and using airborne and terrestrial laser scanning data. We present the contribution of our methodology, which we implemented in an open source software environment using the example project of a 16th century early classical era Ottoman structure, Sinan the Architect’s Şehzade Mosque in Istanbul, Turkey.

  17. Intracellular ROS mediates gas plasma-facilitated cellular transfection in 2D and 3D cultures

    Science.gov (United States)

    Xu, Dehui; Wang, Biqing; Xu, Yujing; Chen, Zeyu; Cui, Qinjie; Yang, Yanjie; Chen, Hailan; Kong, Michael G.

    2016-01-01

    This study reports the potential of cold atmospheric plasma (CAP) as a versatile tool for delivering oligonucleotides into mammalian cells. Compared to lipofection and electroporation methods, plasma transfection showed a better uptake efficiency and less cell death in the transfection of oligonucleotides. We demonstrated that the level of extracellular aqueous reactive oxygen species (ROS) produced by gas plasma is correlated with the uptake efficiency and that this is achieved through an increase of intracellular ROS levels and the resulting increase in cell membrane permeability. This finding was supported by the use of ROS scavengers, which reduced CAP-based uptake efficiency. In addition, we found that cold atmospheric plasma could transfer oligonucleotides such as siRNA and miRNA into cells even in 3D cultures, thus suggesting the potential for unique applications of CAP beyond those provided by standard transfection techniques. Together, our results suggest that cold plasma might provide an efficient technique for the delivery of siRNA and miRNA in 2D and 3D culture models. PMID:27296089

  18. a Semi-Automated Point Cloud Processing Methodology for 3d Cultural Heritage Documentation

    Science.gov (United States)

    Kıvılcım, C. Ö.; Duran, Z.

    2016-06-01

    The preliminary phase in any architectural heritage project is to obtain metric measurements and documentation of the building and its individual elements. On the other hand, conventional measurement techniques require tremendous resources and lengthy project completion times for architectural surveys and 3D model production. Over the past two decades, the widespread use of laser scanning and digital photogrammetry have significantly altered the heritage documentation process. Furthermore, advances in these technologies have enabled robust data collection and reduced user workload for generating various levels of products, from single buildings to expansive cityscapes. More recently, the use of procedural modelling methods and BIM relevant applications for historic building documentation purposes has become an active area of research, however fully automated systems in cultural heritage documentation still remains open. In this paper, we present a semi-automated methodology, for 3D façade modelling of cultural heritage assets based on parametric and procedural modelling techniques and using airborne and terrestrial laser scanning data. We present the contribution of our methodology, which we implemented in an open source software environment using the example project of a 16th century early classical era Ottoman structure, Sinan the Architect's Şehzade Mosque in Istanbul, Turkey.

  19. Multi-cellular 3D human primary liver cell culture elevates metabolic activity under fluidic flow.

    Science.gov (United States)

    Esch, Mandy B; Prot, Jean-Matthieu; Wang, Ying I; Miller, Paula; Llamas-Vidales, Jose Ricardo; Naughton, Brian A; Applegate, Dawn R; Shuler, Michael L

    2015-05-21

    We have developed a low-cost liver cell culture device that creates fluidic flow over a 3D primary liver cell culture that consists of multiple liver cell types, including hepatocytes and non-parenchymal cells (fibroblasts, stellate cells, and Kupffer cells). We tested the performance of the cell culture under fluidic flow for 14 days, finding that hepatocytes produced albumin and urea at elevated levels compared to static cultures. Hepatocytes also responded with induction of P450 (CYP1A1 and CYP3A4) enzyme activity when challenged with P450 inducers, although we did not find significant differences between static and fluidic cultures. Non-parenchymal cells were similarly responsive, producing interleukin 8 (IL-8) when challenged with 10 μM bacterial lipoprotein (LPS). To create the fluidic flow in an inexpensive manner, we used a rocking platform that tilts the cell culture devices at angles between ±12°, resulting in a periodically changing hydrostatic pressure drop between reservoirs and the accompanying periodically changing fluidic flow (average flow rate of 650 μL min(-1), and a maximum shear stress of 0.64 dyne cm(-2)). The increase in metabolic activity is consistent with the hypothesis that, similar to unidirectional fluidic flow, primary liver cell cultures increase their metabolic activity in response to fluidic flow periodically changes direction. Since fluidic flow that changes direction periodically drastically changes the behavior of other cells types that are shear sensitive, our findings support the theory that the increase in hepatic metabolic activity associated with fluidic flow is either activated by mechanisms other than shear sensing (for example increased opportunities for gas and metabolite exchange), or that it follows a shear sensing mechanism that does not depend on the direction of shear. Our mode of device operation allows us to evaluate drugs under fluidic cell culture conditions and at low device manufacturing and operation

  20. Sound localization with head movement: implications for 3-d audio displays.

    Directory of Open Access Journals (Sweden)

    Ken Ian McAnally

    2014-08-01

    Full Text Available Previous studies have shown that the accuracy of sound localization is improved if listeners are allowed to move their heads during signal presentation. This study describes the function relating localization accuracy to the extent of head movement in azimuth. Sounds that are difficult to localize were presented in the free field from sources at a wide range of azimuths and elevations. Sounds remained active until the participants’ heads had rotated through windows ranging in width of 2°, 4°, 8°, 16°, 32°, or 64° of azimuth. Error in determining sound-source elevation and the rate of front/back confusion were found to decrease with increases in azimuth window width. Error in determining sound-source lateral angle was not found to vary with azimuth window width. Implications for 3-d audio displays: The utility of a 3-d audio display for imparting spatial information is likely to be improved if operators are able to move their heads during signal presentation. Head movement may compensate in part for a paucity of spectral cues to sound-source location resulting from limitations in either the audio signals presented or the directional filters (i.e., head-related transfer functions used to generate a display. However, head movements of a moderate size (i.e., through around 32° of azimuth may be required to ensure that spatial information is conveyed with high accuracy.

  1. Assessing the permeability of engineered capillary networks in a 3D culture.

    Directory of Open Access Journals (Sweden)

    Stephanie J Grainger

    Full Text Available Many pathologies are characterized by poor blood vessel growth and reduced nutrient delivery to the surrounding tissue, introducing a need for tissue engineered blood vessels. Our lab has developed a 3D co-culture method to grow interconnected networks of pericyte-invested capillaries, which can anastamose with host vasculature following implantation to restore blood flow to ischemic tissues. However, if the engineered vessels contain endothelial cells (ECs that are misaligned or contain wide junctional gaps, they may function improperly and behave more like the pathologic vessels that nourish tumors. The purpose of this study was to test the resistance to permeability of these networks in vitro, grown with different stromal cell types, as a metric of vessel functionality. A fluorescent dextran tracer was used to visualize transport across the endothelium and the pixel intensity was quantified using a customized MATLAB algorithm. In fibroblast-EC co-cultures, the dextran tracer easily penetrated through the vessel wall and permeability was high through the first 5 days of culture, indicative of vessel immaturity. Beyond day 5, dextran accumulated at the periphery of the vessel, with very little transported across the endothelium. Quantitatively, permeability dropped from initial levels of 61% to 39% after 7 days, and to 7% after 2 weeks. When ECs were co-cultured with bone marrow-derived mesenchymal stem cells (MSCs or adipose-derived stem cells (AdSCs, much tighter control of permeability was achieved. Relative to the EC-fibroblast co-cultures, permeabilities were reduced 41% for the EC-MSC co-cultures and 50% for the EC-AdSC co-cultures after 3 days of culture. By day 14, these permeabilities decreased by 68% and 77% over the EC-fibroblast cultures. Co-cultures containing stem cells exhibit elevated VE-cadherin levels and more prominent EC-EC junctional complexes when compared to cultures containing fibroblasts. These data suggest the stromal

  2. A 3D-psoriatic skin model for dermatological testing: The impact of culture conditions

    Directory of Open Access Journals (Sweden)

    Alexandra Duque-Fernandez

    2016-12-01

    Full Text Available Inadequate representation of the human tissue environment during a preclinical screen can result in inaccurate predictions of compound effects. Consequently, pharmaceutical investigators are searching for preclinical models that closely resemble original tissue for predicting clinical outcomes.The current research aims to compare the impact of using serum-free medium instead of complete culture medium during the last step of psoriatic skin substitute reconstruction. Skin substitutes were produced according to the self-assembly approach.Serum-free conditions have no negative impact on the reconstruction of healthy or psoriatic skin substitutes presented in this study regarding their macroscopic or histological appearances. ATR-FTIR results showed no significant differences in the CH2 bands between psoriatic substitutes cultured with or without serum, thus suggesting that serum deprivation did not have a negative impact on the lipid organization of their stratum corneum. Serum deprivation could even lead to a better organization of healthy skin substitute lipids. Percutaneous analyses demonstrated that psoriatic substitutes cultured in serum-free conditions showed a higher permeability to hydrocortisone compared to controls, while no significant differences in benzoic acid and caffeine penetration profiles were observed.Results obtained with this 3D-psoriatic skin substitute demonstrate the potential and versatility of the model. It could offer good prediction of drug related toxicities at preclinical stages performed in order to avoid unexpected and costly findings in the clinic.Together, these findings offer a new approach for one of the most important challenges of the 21st century, namely, prediction of drug toxicity.•Impact of serum-free conditions during psoriatic skin substitutes reconstruction.•Lipids disorganization of healthy and psoriatic skin substitutes.•Permeation profiles of healthy skin substitutes.•Permeation profiles of

  3. Defining an optimal surface chemistry for pluripotent stem cell culture in 2D and 3D

    Science.gov (United States)

    Zonca, Michael R., Jr.

    Surface chemistry is critical for growing pluripotent stem cells in an undifferentiated state. There is great potential to engineer the surface chemistry at the nanoscale level to regulate stem cell adhesion. However, the challenge is to identify the optimal surface chemistry of the substrata for ES cell attachment and maintenance. Using a high-throughput polymerization and screening platform, a chemically defined, synthetic polymer grafted coating that supports strong attachment and high expansion capacity of pluripotent stem cells has been discovered using mouse embryonic stem (ES) cells as a model system. This optimal substrate, N-[3-(Dimethylamino)propyl] methacrylamide (DMAPMA) that is grafted on 2D synthetic poly(ether sulfone) (PES) membrane, sustains the self-renewal of ES cells (up to 7 passages). DMAPMA supports cell attachment of ES cells through integrin beta1 in a RGD-independent manner and is similar to another recently reported polymer surface. Next, DMAPMA has been able to be transferred to 3D by grafting to synthetic, polymeric, PES fibrous matrices through both photo-induced and plasma-induced polymerization. These 3D modified fibers exhibited higher cell proliferation and greater expression of pluripotency markers of mouse ES cells than 2D PES membranes. Our results indicated that desirable surfaces in 2D can be scaled to 3D and that both surface chemistry and structural dimension strongly influence the growth and differentiation of pluripotent stem cells. Lastly, the feasibility of incorporating DMAPMA into a widely used natural polymer, alginate, has been tested. Novel adhesive alginate hydrogels have been successfully synthesized by either direct polymerization of DMAPMA and methacrylic acid blended with alginate, or photo-induced DMAPMA polymerization on alginate nanofibrous hydrogels. In particular, DMAPMA-coated alginate hydrogels support strong ES cell attachment, exhibiting a concentration dependency of DMAPMA. This research provides a

  4. BioSig3D: High Content Screening of Three-Dimensional Cell Culture Models: e0148379

    National Research Council Canada - National Science Library

    Cemal Cagatay Bilgin; Gerald Fontenay; Qingsu Cheng; Hang Chang; Ju Han; Bahram Parvin

    2016-01-01

    ...) cell culture models that are imaged in full 3D volume. It provides an end-to-end solution for designing high content screening assays, based on colony organization that is derived from segmentation of nuclei in each colony...

  5. Development and Characterization of a Parallelizable Perfusion Bioreactor for 3D Cell Culture

    Science.gov (United States)

    Egger, Dominik; Fischer, Monica; Clementi, Andreas; Ribitsch, Volker; Hansmann, Jan; Kasper, Cornelia

    2017-01-01

    The three dimensional (3D) cultivation of stem cells in dynamic bioreactor systems is essential in the context of regenerative medicine. Still, there is a lack of bioreactor systems that allow the cultivation of multiple independent samples under different conditions while ensuring comprehensive control over the mechanical environment. Therefore, we developed a miniaturized, parallelizable perfusion bioreactor system with two different bioreactor chambers. Pressure sensors were also implemented to determine the permeability of biomaterials which allows us to approximate the shear stress conditions. To characterize the flow velocity and shear stress profile of a porous scaffold in both bioreactor chambers, a computational fluid dynamics analysis was performed. Furthermore, the mixing behavior was characterized by acquisition of the residence time distributions. Finally, the effects of the different flow and shear stress profiles of the bioreactor chambers on osteogenic differentiation of human mesenchymal stem cells were evaluated in a proof of concept study. In conclusion, the data from computational fluid dynamics and shear stress calculations were found to be predictable for relative comparison of the bioreactor geometries, but not for final determination of the optimal flow rate. However, we suggest that the system is beneficial for parallel dynamic cultivation of multiple samples for 3D cell culture processes.

  6. Hepatic stellate cells on poly(DL-lactic acid surfaces control the formation of 3D hepatocyte co-culture aggregates in vitro

    Directory of Open Access Journals (Sweden)

    R J Thomas

    2006-01-01

    Full Text Available Evidence for the functional superiority of cells cultured as 3D aggregates or on 3D scaffolds over conventional 2D monolayer cultures has created interest in material and cell based methods that influence the formation and structure of multicellular aggregates in vitro. We have created a co-culture of primary rat hepatocytes and hepatic stellate cells on a poly(DL-lactic acid surface, a poor substrate for rat hepatocyte adhesion, to study the dynamics of multicellular spheroid formation and the resultant cell arrangement. The poly(DL-lactic acid surface allows dynamic and rapid interaction of hepatocytes and stellate cells to form co-culture spheroids in a complex multistage process (shown by time lapse microscopy. This spheroid morphology supports enhanced cell viability relative to a mono-culture mono-layer system (measured by lactate dehydrogenase leakage. The distribution of the aggregating cell type in the final structure is related to the mechanics of formation i.e. mainly central and peripheral. This study provides a unique and generically applicable insight into the dynamics of multicellular spheroid formation where aggregation is induced by one cell type and imposed on another. This has implications for 3D cell culture models and a wide number of currently used stromal co-culture systems.

  7. 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.

  8. Assessing Drug Efficacy in a Miniaturized Pancreatic Cancer In Vitro 3D Cell Culture Model.

    Science.gov (United States)

    Shelper, Todd B; Lovitt, Carrie J; Avery, Vicky M

    2016-09-01

    Pancreatic cancer continues to have one of the poorest prognoses among all cancers. The drug discovery efforts for this disease have largely failed, with no significant improvement in survival outcomes for advanced pancreatic cancer patients over the past 20 years. Traditional in vitro cell culture techniques have been used extensively in both basic and early drug discovery; however, these systems offer poor models to assess emerging therapeutics. More predictive cell-based models, which better capture the cellular heterogeneity and complexities of solid pancreatic tumors, are urgently needed not only to improve drug discovery success but also to provide insight into the tumor biology. Pancreatic tumors are characterized by a unique micro-environment that is surrounded by a dense stroma. A complex network of interactions between extracellular matrix (ECM) components and the effects of cell-to-cell contacts may enhance survival pathways within in vivo tumors. This biological and physical complexity is lost in traditional cell monolayer models. To explore the predictive potential of a more complex cellular system, a three-dimensional (3D) micro-tumor assay was evaluated. Efficacy of six current chemotherapeutics was determined against a panel of primary and metastatic pancreatic tumor cell lines in a miniaturized ECM-based 3D cell culture system. Suitability for potential use in high-throughput screening applications was assessed, including ascertaining the effects that miniaturization and automation had on assay robustness. Cellular health was determined by utilizing an indirect population-based metabolic activity assay and a direct imaging-based cell viability assay.

  9. Radiation Quality Effects on Transcriptome Profiles in 3-D Cultures After Charged Particle Irradiation

    Science.gov (United States)

    Patel, Zarana S.; Kidane, Yared H.; Huff, Janice L.

    2014-01-01

    In this work, we evaluated the differential effects of low- and high-LET radiation on 3-D organotypic cultures in order to investigate radiation quality impacts on gene expression and cellular responses. Current risk models for assessment of space radiation-induced cancer have large uncertainties because the models for adverse health effects following radiation exposure are founded on epidemiological analyses of human populations exposed to low-LET radiation. Reducing these uncertainties requires new knowledge on the fundamental differences in biological responses (the so-called radiation quality effects) triggered by heavy ion particle radiation versus low-LET radiation associated with Earth-based exposures. In order to better quantify these radiation quality effects in biological systems, we are utilizing novel 3-D organotypic human tissue models for space radiation research. These models hold promise for risk assessment as they provide a format for study of human cells within a realistic tissue framework, thereby bridging the gap between 2-D monolayer culture and animal models for risk extrapolation to humans. To identify biological pathway signatures unique to heavy ion particle exposure, functional gene set enrichment analysis (GSEA) was used with whole transcriptome profiling. GSEA has been used extensively as a method to garner biological information in a variety of model systems but has not been commonly used to analyze radiation effects. It is a powerful approach for assessing the functional significance of radiation quality-dependent changes from datasets where the changes are subtle but broad, and where single gene based analysis using rankings of fold-change may not reveal important biological information.

  10. Polychromatic light-induced osteogenic activity in 2D and 3D cultures.

    Science.gov (United States)

    Ülker, Nazife; Çakmak, Anıl S; Kiremitçi, Arlin S; Gümüşderelioğlu, Menemşe

    2016-11-01

    Photobiomodulation (PBM) has been applied to manipulate cellular responses by using monochromatic light in different wavelengths from ultraviolet (UV) to infrared (IR) region. Until now, an effective wavelength has not been revealed to induce proliferation and/or differentiation of cells. Therefore, in the presented study, we decided to use a specially designed plasma arc light source providing wavelengths between 590 and 1500 nm in order to investigate its biomodulatory effects on chitosan scaffold-supported three-dimensional (3D) cell cultures. For comparison, two-dimensional (2D) cell cultures were also carried out in tissue-culture polystyrene dishes (TCPS). The results showed that light-induced temperature rise did not affect cells when the distance between the light source and the cells was 10 cm and the frequency of administration was daily. Moreover, light was applied for 5 and 10 min to the cells in TCPS and in chitosan scaffold groups, respectively. Cell culture studies under static conditions indicated that polychromatic light significantly stimulated bone nodule formation via the prolonged cell survival and stimulated differentiation of MC3T3-E1 preosteoblastic cells in both TCPS and chitosan scaffold groups. In conclusion, specially designed plasma arc light source used in this study induces formation of bone tissue and so, this light source is proposed as an appropriate system for in vitro bone tissue engineering applications. Statistical analyses were performed with one-way ANOVA by using GraphPad Instat software and standard deviations were calculated by using data of three parallel samples for each group.

  11. Creep Parameterization for the 3-D Mantle: Implications for Geodetic Interpretations of GIA

    Science.gov (United States)

    Ivins, E. R.; Wiens, D.

    2015-12-01

    Providing a volumetrically comprehensive characterization of the variations in the earth's mantle rheology remains one of the grand challenges in geodynamics. An especially difficult task is to make quantitative and non-unique connection between laboratory experiments, seismological mapping and geophysical modeling of glacial isostasy and mantle flow. Advances in seismology and from theoretically modeled high pressure and temperature laboratory experiments make a new set of assessments now possible. We explore the most recent whole mantle seismic models for 3-D shear wave velocity νS( θ, φ, r ): S40RTS (Ritsema et al., 2011) and S362ANI+M (Moulik and Ekström, 2014) together with new data for grain boundary sliding and dislocation mechanics in the olivine/wadsleyite/ringwoodite portions of the upper mantle, connecting these to the well-constrained effective viscosity profiles known for Fennoscandia and Laurentia derived from glacial isostatic adjustment (GIA) data. The latter regions also provide information that connect the operative flow laws for perovskite (Mg,FeSiO3) - periclase (Mg,FeO) mixtures appropriate to levels in the lower mantle corresponding to the high-to-low spin transition occurring at about 2000 km depth (Bardo et al., 2003). Three classic relations for effective viscosity, ηeff, proposed by Ranalli (1990) are employed and evaluated using the two seismic models. We discuss the implications for modeling both Würm-Wisconsin and Little Ice Age deglaciation-related rebound. In addition we discuss the parameter range ηeff, appropriate to modeling GIA in Antarctica, and provide reasonable bounds on errors in forward models. Finally, the implications for GIA removal from GRACE solutions for continent-wide ice sheet mass balance are also treated.

  12. A Novel Flow-Perfusion Bioreactor Supports 3D Dynamic Cell Culture

    Directory of Open Access Journals (Sweden)

    Alexander M. Sailon

    2009-01-01

    Full Text Available Background. Bone engineering requires thicker three-dimensional constructs than the maximum thickness supported by standard cell-culture techniques (2 mm. A flow-perfusion bioreactor was developed to provide chemotransportation to thick (6 mm scaffolds. Methods. Polyurethane scaffolds, seeded with murine preosteoblasts, were loaded into a novel bioreactor. Control scaffolds remained in static culture. Samples were harvested at days 2, 4, 6, and 8 and analyzed for cellular distribution, viability, metabolic activity, and density at the periphery and core. Results. By day 8, static scaffolds had a periphery cell density of 67%±5.0%, while in the core it was 0.3%±0.3%. Flow-perfused scaffolds demonstrated peripheral cell density of 94%±8.3% and core density of 76%±3.1% at day 8. Conclusions. Flow perfusion provides chemotransportation to thick scaffolds. This system may permit high throughput study of 3D tissues in vitro and enable prefabrication of biological constructs large enough to solve clinical problems.

  13. Engineering a 3D microfluidic culture platform for tumor-treating field application

    Science.gov (United States)

    Pavesi, Andrea; Adriani, Giulia; Tay, Andy; Warkiani, Majid Ebrahimi; Yeap, Wei Hseun; Wong, Siew Cheng; Kamm, Roger D.

    2016-05-01

    The limitations of current cancer therapies highlight the urgent need for a more effective therapeutic strategy. One promising approach uses an alternating electric field; however, the mechanisms involved in the disruption of the cancer cell cycle as well as the potential adverse effects on non-cancerous cells must be clarified. In this study, we present a novel microfluidic device with embedded electrodes that enables the application of an alternating electric field therapy to cancer cells in a 3D extracellular matrix. To demonstrate the potential of our system to aid in designing and testing new therapeutic approaches, cancer cells and cancer cell aggregates were cultured individually or co-cultured with endothelial cells. The metastatic potential of the cancer cells was reduced after electric field treatment. Moreover, the proliferation rate of the treated cancer cells was lower compared with that of the untreated cells, whereas the morphologies and proliferative capacities of the endothelial cells were not significantly affected. These results demonstrate that our novel system can be used to rapidly screen the effect of an alternating electric field on cancer and normal cells within an in vivo-like microenvironment with the potential to optimize treatment protocols and evaluate synergies between tumor-treating field treatment and chemotherapy.

  14. The cultural divide: exponential growth in classical 2D and metabolic equilibrium in 3D environments.

    Directory of Open Access Journals (Sweden)

    Krzysztof Wrzesinski

    Full Text Available INTRODUCTION: Cellular metabolism can be considered to have two extremes: one is characterized by exponential growth (in 2D cultures and the other by a dynamic equilibrium (in 3D cultures. We have analyzed the proteome and cellular architecture at these two extremes and found that they are dramatically different. RESULTS: Structurally, actin organization is changed, microtubules are increased and keratins 8 and 18 decreased. Metabolically, glycolysis, fatty acid metabolism and the pentose phosphate shunt are increased while TCA cycle and oxidative phosphorylation is unchanged. Enzymes involved in cholesterol and urea synthesis are increased consistent with the attainment of cholesterol and urea production rates seen in vivo. DNA repair enzymes are increased even though cells are predominantly in Go. Transport around the cell--along the microtubules, through the nuclear pore and in various types of vesicles has been prioritized. There are numerous coherent changes in transcription, splicing, translation, protein folding and degradation. The amount of individual proteins within complexes is shown to be highly coordinated. Typically subunits which initiate a particular function are present in increased amounts compared to other subunits of the same complex. SUMMARY: We have previously demonstrated that cells at dynamic equilibrium can match the physiological performance of cells in tissues in vivo. Here we describe the multitude of protein changes necessary to achieve this performance.

  15. Scaffolds for 3D in vitro culture of neural lineage cells.

    Science.gov (United States)

    Murphy, Ashley R; Laslett, Andrew; O'Brien, Carmel M; Cameron, Neil R

    2017-03-01

    Understanding how neurodegenerative disorders develop is not only a key challenge for researchers but also for the wider society, given the rapidly aging populations in developed countries. Advances in this field require new tools with which to recreate neural tissue in vitro and produce realistic disease models. This in turn requires robust and reliable systems for performing 3D in vitro culture of neural lineage cells. This review provides a state of the art update on three-dimensional culture systems for in vitro development of neural tissue, employing a wide range of scaffold types including hydrogels, solid porous polymers, fibrous materials and decellularised tissues as well as microfluidic devices and lab-on-a-chip systems. To provide some context with in vivo development of the central nervous system (CNS), we also provide a brief overview of the neural stem cell niche, neural development and neural differentiation in vitro. We conclude with a discussion of future directions for this exciting and important field of biomaterials research.

  16. UAV and Computer Vision in 3D Modeling of Cultural Heritage in Southern Italy

    Science.gov (United States)

    Barrile, Vincenzo; Gelsomino, Vincenzo; Bilotta, Giuliana

    2017-08-01

    On the Waterfront Italo Falcomatà of Reggio Calabria you can admire the most extensive tract of the walls of the Hellenistic period of ancient city of Rhegion. The so-called Greek Walls are one of the most significant and visible traces of the past linked to the culture of Ancient Greece in the site of Reggio Calabria territory. Over the years this stretch of wall has always been a part, to the reconstruction of Reggio after the earthquake of 1783, the outer walls at all times, restored countless times, to cope with the degradation of the time and the adjustments to the technical increasingly innovative and sophisticated siege. They were the subject of several studies on history, for the study of the construction techniques and the maintenance and restoration of the same. This note describes the methodology for the implementation of a three-dimensional model of the Greek Walls conducted by the Geomatics Laboratory, belonging to DICEAM Department of University “Mediterranea” of Reggio Calabria. 3D modeling we made is based on imaging techniques, such as Digital Photogrammetry and Computer Vision, by using a drone. The acquired digital images were then processed using commercial software Agisoft PhotoScan. The results denote the goodness of the technique used in the field of cultural heritage, attractive alternative to more expensive and demanding techniques such as laser scanning.

  17. 3D Simulations of Helmet Streamer Dynamics and Implications for the Slow Solar Wind

    Science.gov (United States)

    Higginson, Aleida K.; Antiochos, Spiro K.; DeVore, C. R.; Zurbuchen, Thomas H.

    2015-04-01

    The source of the slow solar wind at the Sun is still an issue of intense debate in solar and heliospheric physics. Because the majority of the solar wind observed at Earth is slow wind, understanding its origin is essential for understanding and predicting Earth’s space weather environment. In-situ and remote observations show that, when compared to the fast wind, the slow solar wind corresponds to higher freeze-in temperatures, as indicated by charge-state ratios, and more corona-like elemental abundance ratios. These results indicate that the most likely source for the slow wind is the hot plasma in the closed-field corona, but the release mechanism(s) for the wind from the closed-field regions is far from understood. We perform fully dynamic, 3D MHD simulations in order to the study the opening and closing of the Sun’s magnetic field that leads to the escape of the slow solar wind. In particular, we calculate the dynamics of helmet streamers that are driven by photospheric motions such as supergranular flows. We determine in detail the opening and closing of coronal flux, and discuss the implications of our results for theories of slow wind origin, especially the S-Web model. We also determine observational signatures for the upcoming inner heliosphere missions Solar Orbiter and Solar Probe Plus.This work was supported by the NASA SR&T and TR&T Programs.

  18. Application of 3D hydrodynamic and particle tracking models for better environmental management of finfish culture

    Science.gov (United States)

    Moreno Navas, Juan; Telfer, Trevor C.; Ross, Lindsay G.

    2011-04-01

    Hydrographic conditions, and particularly current speeds, have a strong influence on the management of fish cage culture. These hydrodynamic conditions can be used to predict particle movement within the water column and the results used to optimise environmental conditions for effective site selection, setting of environmental quality standards, waste dispersion, and potential disease transfer. To this end, a 3D hydrodynamic model, MOHID, has been coupled to a particle tracking model to study the effects of mean current speed, quiescent water periods and bulk water circulation in Mulroy Bay, Co. Donegal Ireland, an Irish fjard (shallow fjordic system) important to the aquaculture industry. A Lagangrian method simulated the instantaneous release of "particles" emulating discharge from finfish cages to show the behaviour of waste in terms of water circulation and water exchange. The 3D spatial models were used to identify areas of mixed and stratified water using a version of the Simpson-Hunter criteria, and to use this in conjunction with models of current flow for appropriate site selection for salmon aquaculture. The modelled outcomes for stratification were in good agreement with the direct measurements of water column stratification based on observed density profiles. Calculations of the Simpson-Hunter tidal parameter indicated that most of Mulroy Bay was potentially stratified with a well mixed region over the shallow channels where the water is faster flowing. The fjard was characterised by areas of both very low and high mean current speeds, with some areas having long periods of quiescent water. The residual current and the particle tracking animations created through the models revealed an anticlockwise eddy that may influence waste dispersion and potential for disease transfer, among salmon cages and which ensures that the retention time of waste substances from cages is extended. The hydrodynamic model results were incorporated into the ArcView TM GIS

  19. Radiation Quality Effects on Transcriptome Profiles in 3-d Cultures After Particle Irradiation

    Science.gov (United States)

    Patel, Z. S.; Kidane, Y. H.; Huff, J. L.

    2014-01-01

    In this work, we evaluate the differential effects of low- and high-LET radiation on 3-D organotypic cultures in order to investigate radiation quality impacts on gene expression and cellular responses. Reducing uncertainties in current risk models requires new knowledge on the fundamental differences in biological responses (the so-called radiation quality effects) triggered by heavy ion particle radiation versus low-LET radiation associated with Earth-based exposures. We are utilizing novel 3-D organotypic human tissue models that provide a format for study of human cells within a realistic tissue framework, thereby bridging the gap between 2-D monolayer culture and animal models for risk extrapolation to humans. To identify biological pathway signatures unique to heavy ion particle exposure, functional gene set enrichment analysis (GSEA) was used with whole transcriptome profiling. GSEA has been used extensively as a method to garner biological information in a variety of model systems but has not been commonly used to analyze radiation effects. It is a powerful approach for assessing the functional significance of radiation quality-dependent changes from datasets where the changes are subtle but broad, and where single gene based analysis using rankings of fold-change may not reveal important biological information. We identified 45 statistically significant gene sets at 0.05 q-value cutoff, including 14 gene sets common to gamma and titanium irradiation, 19 gene sets specific to gamma irradiation, and 12 titanium-specific gene sets. Common gene sets largely align with DNA damage, cell cycle, early immune response, and inflammatory cytokine pathway activation. The top gene set enriched for the gamma- and titanium-irradiated samples involved KRAS pathway activation and genes activated in TNF-treated cells, respectively. Another difference noted for the high-LET samples was an apparent enrichment in gene sets involved in cycle cycle/mitotic control. It is

  20. Differences in growth properties of endometrial cancer in three dimensional (3D) culture and 2D cell monolayer

    Energy Technology Data Exchange (ETDEWEB)

    Chitcholtan, Kenny, E-mail: kenny.chitcholtan@otago.ac.nz [Department of Obstetrics and Gynaecology, University of Otago, Christchurch, 2 Riccarton Avenue, Christchurch 8011 (New Zealand); Asselin, Eric, E-mail: Eric.Asselin@uqtr.ca [Department of Chemistry and Biology, University of Quebec, at Trois-Rivières, C.P. 500, Trois-Rivières, Quebec, Canada G9A 5H7 (Canada); Parent, Sophie, E-mail: Sophie.Parent@uqtr.ca [Department of Chemistry and Biology, University of Quebec, at Trois-Rivières, C.P. 500, Trois-Rivières, Quebec, Canada G9A 5H7 (Canada); Sykes, Peter H., E-mail: peter.sykes@otago.ac.nz [Department of Obstetrics and Gynaecology, University of Otago, Christchurch, 2 Riccarton Avenue, Christchurch 8011 (New Zealand); Evans, John J., E-mail: john.evans@otago.ac.nz [Department of Obstetrics and Gynaecology, University of Otago, Christchurch, 2 Riccarton Avenue, Christchurch 8011 (New Zealand); Centre of Neuroendocrinology and The MacDiarmid Institute of Advanced Materials and Nanotechnology, University of Otago, Christchurch, 2 Riccarton Avenue, Christchurch 8011 (New Zealand)

    2013-01-01

    Three-dimensional (3D) in vitro models have an invaluable role in understanding the behaviour of tumour cells in a well defined microenvironment. This is because some aspects of tumour characteristics cannot be fully recapitulated in a cell monolayer (2D). In the present study, we compared growth patterns, expression of signalling molecules, and metabolism-associated proteins of endometrial cancer cell lines in 3D and 2D cell cultures. Cancer cells formed spherical structures in 3D reconstituted basement membrane (3D rBM), and the morphological appearance was cell line dependent. Cell differentiation was observed after 8 days in the 3D rBM. There was reduced proliferation, detected by less expression of PCNA in 3D rBM than in 2D cell monolayers. The addition of exogenous epidermal growth factor (EGF) to cancer cells induced phosphorylation of EGFR and Akt in both cell culture conditions. The uptake of glucose was selectively altered in the 3D rBM, but there was a lack of association with Glut-1 expression. The secretion of vascular endothelial growth factor (VEGF) and prostaglandin E{sub 2} (PGE{sub 2}) was selectively altered in 3D rBM, and it was cell line dependent. Our data demonstrated that 3D rBM as an in vitro model can influence proliferation and metabolism of endometrial cancer cell behaviour compared to 2D cell monolayer. Changes are specific to individual cell types. The use of 3D rBM is, therefore, important in the in vitro study of targeted anticancer therapies.

  1. Culture phases, cytotoxicity and protein expressions of agarose hydrogel induced Sp2/0, A549, MCF-7 cell line 3D cultures.

    Science.gov (United States)

    Ravi, Maddaly; Kaviya, S R; Paramesh, V

    2016-05-01

    Advancements in cell cultures are occurring at a rapid pace, an important direction is culturing cells in 3D conditions. We demonstrate the usefulness of agarose hydrogels in obtaining 3 dimensional aggregates of three cell lines, A549, MCF-7 and Sp2/0. The differences in culture phases, susceptibility to cisplatin-induced cytotoxicity are studied. Also, the 3D aggregates of the three cell lines were reverted into 2D cultures and the protein profile differences among the 2D, 3D and revert cultures were studied. The analysis of protein profile differences using UniProt data base further augment the usefulness of agarose hydrogels for obtaining 3D cell cultures.

  2. Understanding the impact of 2D and 3D fibroblast cultures on in vitro breast cancer models.

    Directory of Open Access Journals (Sweden)

    Kyung Eun Sung

    Full Text Available The utilization of 3D, physiologically relevant in vitro cancer models to investigate complex interactions between tumor and stroma has been increasing. Prior work has generally focused on the cancer cells and, the role of fibroblast culture conditions on tumor-stromal cell interactions is still largely unknown. Here, we focus on the stroma by comparing functional behaviors of human mammary fibroblasts (HMFs cultured in 2D and 3D and their effects on the invasive progression of breast cancer cells (MCF10DCIS.com. We identified increased levels of several paracrine factors from HMFs cultured in 3D conditions that drive the invasive transition. Using a microscale co-culture model with improved compartmentalization and sensitivity, we demonstrated that HMFs cultured in 3D intensify the promotion of the invasive progression through the HGF/c-Met interaction. This study highlights the importance of the 3D stromal microenvironment in the development of multiple cell type in vitro cancer models.

  3. Critical analysis of 3-D organoid in vitro cell culture models for high-throughput drug candidate toxicity assessments.

    Science.gov (United States)

    Astashkina, Anna; Grainger, David W

    2014-04-01

    Drug failure due to toxicity indicators remains among the primary reasons for staggering drug attrition rates during clinical studies and post-marketing surveillance. Broader validation and use of next-generation 3-D improved cell culture models are expected to improve predictive power and effectiveness of drug toxicological predictions. However, after decades of promising research significant gaps remain in our collective ability to extract quality human toxicity information from in vitro data using 3-D cell and tissue models. Issues, challenges and future directions for the field to improve drug assay predictive power and reliability of 3-D models are reviewed. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. Chemotherapeutic efficiency of drugs in vitro: Comparison of doxorubicin exposure in 3D and 2D culture matrices.

    Science.gov (United States)

    Casey, A; Gargotti, M; Bonnier, F; Byrne, H J

    2016-06-01

    The interest in the use of 3D matrices for in vitro analysis, with a view to increasing the relevance of in vitro studies and reducing the dependence on in vivo studies, has been growing in recent years. Cells grown in a 3D in vitro matrix environment have been reported to exhibit significantly different properties to those in a conventional 2D culture environment. However, comparison of 2D and 3D cell culture models have recently been noted to result in differing responses of cytotoxic assays, without any associated change in viability. The effect was attributed to differing conversion rates and effective concentrations of the resazurin assay in 2D and 3D environments, rather than differences in cellular metabolism. In this study, the efficacy of a chemotherapeutic agent, doxorubicin, is monitored and compared in conventional 2D and 3D collagen gel exposures of immortalized human cervical cells. Viability was monitored with the aid of the Alamar Blue assay and drug internalisation was verified using confocal microscopy. Drug uptake and retention within the collagen matrix was monitored by absorption spectroscopy. The viability studies showed apparent differences between the 2D and 3D culture systems, the differences attributed in part to the physical transition from 2D to a 3D environment causing alterations to dye resazurin uptake and conversion rates. The use of 3D culture matrices has widely been interpreted to result in "reduced" toxicity or cellular "resistance" to the chemotherapeutic agent. The results of this study show that the reduced efficiency of the drug to cells grown in the 3D environment can be accounted for by a sequential reduction of the effective concentration of the test compound and assay. This is due to absorption within the collagen gel inducing a higher uptake of both drug and assay thereby influencing the toxic impact of the drug and conversion rate of resazurin, and. The increased effective surface area of the cell exposed to the drug

  5. Bioinspired Tuning of Hydrogel Permeability-Rigidity Dependency for 3D Cell Culture

    Science.gov (United States)

    Lee, Min Kyung; Rich, Max H.; Baek, Kwanghyun; Lee, Jonghwi; Kong, Hyunjoon

    2015-03-01

    Hydrogels are being extensively used for three-dimensional immobilization and culture of cells in fundamental biological studies, biochemical processes, and clinical treatments. However, it is still a challenge to support viability and regulate phenotypic activities of cells in a structurally stable gel, because the gel becomes less permeable with increasing rigidity. To resolve this challenge, this study demonstrates a unique method to enhance the permeability of a cell-laden hydrogel while avoiding a significant change in rigidity of the gel. Inspired by the grooved skin textures of marine organisms, a hydrogel is assembled to present computationally optimized micro-sized grooves on the surface. Separately, a gel is engineered to preset aligned microchannels similar to a plant's vascular bundles through a uniaxial freeze-drying process. The resulting gel displays significantly increased water diffusivity with reduced changes of gel stiffness, exclusively when the microgrooves and microchannels are aligned together. No significant enhancement of rehydration is achieved when the microgrooves and microchannels are not aligned. Such material design greatly enhances viability and neural differentiation of stem cells and 3D neural network formation within the gel.

  6. Self-Organization of Polarized Cerebellar Tissue in 3D Culture of Human Pluripotent Stem Cells

    Directory of Open Access Journals (Sweden)

    Keiko Muguruma

    2015-02-01

    Full Text Available During cerebellar development, the main portion of the cerebellar plate neuroepithelium gives birth to Purkinje cells and interneurons, whereas the rhombic lip, the germinal zone at its dorsal edge, generates granule cells and cerebellar nuclei neurons. However, it remains elusive how these components cooperate to form the intricate cerebellar structure. Here, we found that a polarized cerebellar structure self-organizes in 3D human embryonic stem cell (ESC culture. The self-organized neuroepithelium differentiates into electrophysiologically functional Purkinje cells. The addition of fibroblast growth factor 19 (FGF19 promotes spontaneous generation of dorsoventrally polarized neural-tube-like structures at the level of the cerebellum. Furthermore, addition of SDF1 and FGF19 promotes the generation of a continuous cerebellar plate neuroepithelium with rhombic-lip-like structure at one end and a three-layer cytoarchitecture similar to the embryonic cerebellum. Thus, human-ESC-derived cerebellar progenitors exhibit substantial self-organizing potential for generating a polarized structure reminiscent of the early human cerebellum at the first trimester.

  7. Dynamic, large-scale profiling of transcription factor activity from live cells in 3D culture.

    Directory of Open Access Journals (Sweden)

    Michael S Weiss

    Full Text Available BACKGROUND: Extracellular activation of signal transduction pathways and their downstream target transcription factors (TFs are critical regulators of cellular processes and tissue development. The intracellular signaling network is complex, and techniques that quantify the activities of numerous pathways and connect their activities to the resulting phenotype would identify the signals and mechanisms regulating tissue development. The ability to investigate tissue development should capture the dynamic pathway activity and requires an environment that supports cellular organization into structures that mimic in vivo phenotypes. Taken together, our objective was to develop cellular arrays for dynamic, large-scale quantification of TF activity as cells organized into spherical structures within 3D culture. METHODOLOGY/PRINCIPAL FINDINGS: TF-specific and normalization reporter constructs were delivered in parallel to a cellular array containing a well-established breast cancer cell line cultured in Matrigel. Bioluminescence imaging provided a rapid, non-invasive, and sensitive method to quantify luciferase levels, and was applied repeatedly on each sample to monitor dynamic activity. Arrays measuring 28 TFs identified up to 19 active, with 13 factors changing significantly over time. Stimulation of cells with β-estradiol or activin A resulted in differential TF activity profiles evolving from initial stimulation of the ligand. Many TFs changed as expected based on previous reports, yet arrays were able to replicate these results in a single experiment. Additionally, arrays identified TFs that had not previously been linked with activin A. CONCLUSIONS/SIGNIFICANCE: This system provides a method for large-scale, non-invasive, and dynamic quantification of signaling pathway activity as cells organize into structures. The arrays may find utility for investigating mechanisms regulating normal and abnormal tissue growth, biomaterial design, or as a

  8. AlgiMatrix™ based 3D cell culture system as an in-vitro tumor model for anticancer studies.

    Directory of Open Access Journals (Sweden)

    Chandraiah Godugu

    Full Text Available BACKGROUND: Three-dimensional (3D in-vitro cultures are recognized for recapitulating the physiological microenvironment and exhibiting high concordance with in-vivo conditions. Taking the advantages of 3D culture, we have developed the in-vitro tumor model for anticancer drug screening. METHODS: Cancer cells grown in 6 and 96 well AlgiMatrix™ scaffolds resulted in the formation of multicellular spheroids in the size range of 100-300 µm. Spheroids were grown in two weeks in cultures without compromising the growth characteristics. Different marketed anticancer drugs were screened by incubating them for 24 h at 7, 9 and 11 days in 3D cultures and cytotoxicity was measured by AlamarBlue® assay. Effectiveness of anticancer drug treatments were measured based on spheroid number and size distribution. Evaluation of apoptotic and anti-apoptotic markers was done by immunohistochemistry and RT-PCR. The 3D results were compared with the conventional 2D monolayer cultures. Cellular uptake studies for drug (Doxorubicin and nanoparticle (NLC were done using spheroids. RESULTS: IC(50 values for anticancer drugs were significantly higher in AlgiMatrix™ systems compared to 2D culture models. The cleaved caspase-3 expression was significantly decreased (2.09 and 2.47 folds respectively for 5-Fluorouracil and Camptothecin in H460 spheroid cultures compared to 2D culture system. The cytotoxicity, spheroid size distribution, immunohistochemistry, RT-PCR and nanoparticle penetration data suggested that in vitro tumor models show higher resistance to anticancer drugs and supporting the fact that 3D culture is a better model for the cytotoxic evaluation of anticancer drugs in vitro. CONCLUSION: The results from our studies are useful to develop a high throughput in vitro tumor model to study the effect of various anticancer agents and various molecular pathways affected by the anticancer drugs and formulations.

  9. Visualization of Mesenchymal Stromal Cells in 2Dand 3D-Cultures by Scanning Electron Microscopy with Lanthanide Contrasting.

    Science.gov (United States)

    Novikov, I A; Vakhrushev, I V; Antonov, E N; Yarygin, K N; Subbot, A M

    2017-02-01

    Mesenchymal stromal cells from deciduous teeth in 2D- and 3D-cultures on culture plastic, silicate glass, porous polystyrene, and experimental polylactoglycolide matrices were visualized by scanning electron microscopy with lanthanide contrasting. Supravital staining of cell cultures with a lanthanide-based dye (neodymium chloride) preserved normal cell morphology and allowed assessment of the matrix properties of the carriers. The developed approach can be used for the development of biomaterials for tissue engineering.

  10. Inkjet4Tex: Creative implications of 3D inkjet printing technologies for textiles

    OpenAIRE

    Campbell, J.R.

    2009-01-01

    This project expands future applied-design capabilities for textiles as a function of inkjet deposition technology. The project investigates 3D inkjet rapid-production tools’ potential, focusing on creative gaps in the developing technology in its application to the textile design process. As such, the research investigates future design possibilities for inkjet printing technology in the creation of 3D textile structures and surfaces. The research “demonstrates how tacit knowledge can be emp...

  11. 3D Airflow patterns over coastal foredunes: implications for aeolian sediment transport

    Science.gov (United States)

    Jackson, Derek W. T.; Cooper, Andrew G.; Baas, Andreas C. W.; Lynch, Kevin; Beyers, Meiring

    2010-05-01

    A fundamental criterion for the development of coastal sand dunes is usually highlighted as a significant onshore wind component of the local wind field. The presence of large sand dune systems on coasts where the predominant wind blows offshore is therefore difficult to explain and usually they are attributed to the past occurrence of onshore winds and, by implication, subsequent changes in climate. Recent studies have shown that offshore winds can be deflected or 'steered' by existing dunes so that their direction changes. This can occur to such an extent that a process known as 'flow reversal' can arise, whereby the initially offshore wind actually flows onshore at the beach. This process is important because it can cause sand to be blown from the beach and into the dunes, causing them to grow. This may be central in explaining the presence of extensive dunes on coasts where the dominant wind is offshore, but is also important in how dunes recover after periods of wave erosion during storms. Offshore winds have traditionally been excluded from sediment budget calculations for coastal dunes, but when they do transport sand onshore, this may have been an important oversight leading to significant underestimates of the volume of sand being transported by wind. This work investigates the controls on the processes and the mechanisms involved in deformation of the flow and resulting sediment transport at coastal foredunes in Northern Ireland. We use a combination of field measurement of wind and sediment transport coupled with state-of-the-art aerodynamic modelling using computational fluid dynamics (CFD) and 3-D sonic anemometry. Our working hypothesis is that offshore winds contribute substantially to foredune behaviour on leeside coasts. Preliminary results show strong reverse flow eddies in the seaward side of the foredunes during offshore wind events. These secondary flow reversals have been above velocity threshold and are transport capable. Using CFD modelling

  12. Potential and limitations of microscopy and Raman spectroscopy for live-cell analysis of 3D cell cultures.

    Science.gov (United States)

    Charwat, Verena; Schütze, Karin; Holnthoner, Wolfgang; Lavrentieva, Antonina; Gangnus, Rainer; Hofbauer, Pablo; Hoffmann, Claudia; Angres, Brigitte; Kasper, Cornelia

    2015-07-10

    Today highly complex 3D cell culture formats that closely mimic the in vivo situation are increasingly available. Despite their wide use, the development of analytical methods and tools that can work within the depth of 3D-tissue constructs lags behind. In order to get the most information from a 3D cell sample, adequate and reliable assays are required. However, the majority of tools and methods used today have been originally designed for 2D cell cultures and translation to a 3D environment is in general not trivial. Ideally, an analytical method should be non-invasive and allow for repeated observation of living cells in order to detect dynamic changes in individual cells within the 3D cell culture. Although well-established laser confocal microscopy can be used for these purposes, this technique has serious limitations including penetration depth and availability. Focusing on two relevant analytical methods for live-cell monitoring, we discuss the current challenges of analyzing living 3D samples: microscopy, which is the most widely used technology to observe and examine cell cultures, has been successfully adapted for 3D samples by recording of so-called "z-stacks". However the required equipment is generally very expensive and therefore access is often limited. Consequently alternative and less advanced approaches are often applied that cannot capture the full structural complexity of a 3D sample. Similarly, image analysis tools for quantification of microscopic images range from highly specialized and costly to simplified and inexpensive. Depending on the actual sample composition and scientific question the best approach needs to be assessed individually. Another more recently introduced technology for non-invasive cell analysis is Raman micro-spectroscopy. It enables label-free identification of cellular metabolic changes with high sensitivity and has already been successful applied to 2D and 3D cell cultures. However, its future significance for cell

  13. A 3D Sphere Culture System Containing Functional Polymers for Large-Scale Human Pluripotent Stem Cell Production

    Directory of Open Access Journals (Sweden)

    Tomomi G. Otsuji

    2014-05-01

    Full Text Available Utilizing human pluripotent stem cells (hPSCs in cell-based therapy and drug discovery requires large-scale cell production. However, scaling up conventional adherent cultures presents challenges of maintaining a uniform high quality at low cost. In this regard, suspension cultures are a viable alternative, because they are scalable and do not require adhesion surfaces. 3D culture systems such as bioreactors can be exploited for large-scale production. However, the limitations of current suspension culture methods include spontaneous fusion between cell aggregates and suboptimal passaging methods by dissociation and reaggregation. 3D culture systems that dynamically stir carrier beads or cell aggregates should be refined to reduce shearing forces that damage hPSCs. Here, we report a simple 3D sphere culture system that incorporates mechanical passaging and functional polymers. This setup resolves major problems associated with suspension culture methods and dynamic stirring systems and may be optimal for applications involving large-scale hPSC production.

  14. Getting it right: 3D cell cultures for the assessment of photosensitizers for photodynamic therapy

    OpenAIRE

    SENGE, MATHIAS

    2015-01-01

    PUBLISHED Core quote “Cancer drug screening is rapidly moving towards the use of 3D cell models. The added complexity of PDT action makes this a conditio sine qua non for the screening of new photosensitizers.”

  15. 3D Documentation and BIM Modeling of Cultural Heritage Structures Using UAVs: The Case of the Foinikaria Church

    Science.gov (United States)

    Themistocleous, K.; Agapiou, A.; Hadjimitsis, D.

    2016-10-01

    The documentation of architectural cultural heritage sites has traditionally been expensive and labor-intensive. New innovative technologies, such as Unmanned Aerial Vehicles (UAVs), provide an affordable, reliable and straightforward method of capturing cultural heritage sites, thereby providing a more efficient and sustainable approach to documentation of cultural heritage structures. In this study, hundreds of images of the Panagia Chryseleousa church in Foinikaria, Cyprus were taken using a UAV with an attached high resolution camera. The images were processed to generate an accurate digital 3D model by using Structure in Motion techniques. Building Information Model (BIM) was then used to generate drawings of the church. The methodology described in the paper provides an accurate, simple and cost-effective method of documenting cultural heritage sites and generating digital 3D models using novel techniques and innovative methods.

  16. 5D Modelling: An Efficient Approach for Creating Spatiotemporal Predictive 3D Maps of Large-Scale Cultural Resources

    Science.gov (United States)

    Doulamis, A.; Doulamis, N.; Ioannidis, C.; Chrysouli, C.; Grammalidis, N.; Dimitropoulos, K.; Potsiou, C.; Stathopoulou, E.-K.; Ioannides, M.

    2015-08-01

    Outdoor large-scale cultural sites are mostly sensitive to environmental, natural and human made factors, implying an imminent need for a spatio-temporal assessment to identify regions of potential cultural interest (material degradation, structuring, conservation). On the other hand, in Cultural Heritage research quite different actors are involved (archaeologists, curators, conservators, simple users) each of diverse needs. All these statements advocate that a 5D modelling (3D geometry plus time plus levels of details) is ideally required for preservation and assessment of outdoor large scale cultural sites, which is currently implemented as a simple aggregation of 3D digital models at different time and levels of details. The main bottleneck of such an approach is its complexity, making 5D modelling impossible to be validated in real life conditions. In this paper, a cost effective and affordable framework for 5D modelling is proposed based on a spatial-temporal dependent aggregation of 3D digital models, by incorporating a predictive assessment procedure to indicate which regions (surfaces) of an object should be reconstructed at higher levels of details at next time instances and which at lower ones. In this way, dynamic change history maps are created, indicating spatial probabilities of regions needed further 3D modelling at forthcoming instances. Using these maps, predictive assessment can be made, that is, to localize surfaces within the objects where a high accuracy reconstruction process needs to be activated at the forthcoming time instances. The proposed 5D Digital Cultural Heritage Model (5D-DCHM) is implemented using open interoperable standards based on the CityGML framework, which also allows the description of additional semantic metadata information. Visualization aspects are also supported to allow easy manipulation, interaction and representation of the 5D-DCHM geometry and the respective semantic information. The open source 3DCity

  17. Main Features of a 3d GIS for a Monumental Complex with AN Historical-Cultural Relevance

    Science.gov (United States)

    Scianna, A.; La Guardia, M.

    2017-05-01

    The last achievements of technologies in geomatics especially in survey and restitution of 3D models (UAV/drones and laser scanner technologies) generated new procedures and higher standards of quality in representation of archaeological sites. Together with Geomatics, the recent development of Information and Communication Technologies (ICT) strongly contribute to document and the Cultural Heritage (CH). The representation and documentation of CH using these new technologies has became necessary in order to satisfy different needs: - for restorers in order to acquire a deep knowledge of the cultural good and to define possible strategies of restoration; - for the conservation of information, allowing to preserve the 3D geometry of the monumental complex with the integration of descriptions about architectural elements; - for touristic aims, giving the opportunity of sharing CH information on web, allowing users to visit and explore, in a virtual way, monumental complexes, acquiring information details about architectural elements or the history of monumental complex. Looking through these new scenarios, the development of a 3D Geographic Information System (GIS) applied to a cultural good could be, today, an added value of fundamental importance for full description and data management of monumental complexes. In this work, the main features necessary for the correct construction of a 3D GIS of a monumental complex will be analyzed, with a particular focus on the possibilities for creating a standardized procedure to follow.

  18. Alteration of cellular behavior and response to PI3K pathway inhibition by culture in 3D collagen gels.

    Directory of Open Access Journals (Sweden)

    Brian Fallica

    Full Text Available Most investigations into cancer cell drug response are performed with cells cultured on flat (2D tissue culture plastic. Emerging research has shown that the presence of a three-dimensional (3D extracellular matrix (ECM is critical for normal cell behavior including migration, adhesion, signaling, proliferation and apoptosis. In this study we investigate differences between cancer cell signaling in 2D culture and a 3D ECM, employing real-time, live cell tracking to directly observe U2OS human osteosarcoma and MCF7 human breast cancer cells embedded in type 1 collagen gels. The activation of the important PI3K signaling pathway under these different growth conditions is studied, and the response to inhibition of both PI3K and mTOR with PI103 investigated. Cells grown in 3D gels show reduced proliferation and migration as well as reduced PI3K pathway activation when compared to cells grown in 2D. Our results quantitatively demonstrate that a collagen ECM can protect U2OS cells from PI103. Overall, our data suggests that 3D gels may provide a better medium for investigation of anti-cancer drugs than 2D monolayers, therefore allowing better understanding of cellular response and behavior in native like environments.

  19. A spheroid-based 3-D culture model for pancreatic cancer drug testing, using the acid phosphatase assay

    Energy Technology Data Exchange (ETDEWEB)

    Wen, Z.; Liao, Q.; Hu, Y.; You, L.; Zhou, L.; Zhao, Y. [Department of General Surgery, Peking Union Medical College Hospital, Chinese Academy of Medical Science and Peking Union Medical College, Tsinghua University, Beijing (China)

    2013-08-10

    Current therapy for pancreatic cancer is multimodal, involving surgery and chemotherapy. However, development of pancreatic cancer therapies requires a thorough evaluation of drug efficacy in vitro before animal testing and subsequent clinical trials. Compared to two-dimensional culture of cell monolayer, three-dimensional (3-D) models more closely mimic native tissues, since the tumor microenvironment established in 3-D models often plays a significant role in cancer progression and cellular responses to the drugs. Accumulating evidence has highlighted the benefits of 3-D in vitro models of various cancers. In the present study, we have developed a spheroid-based, 3-D culture of pancreatic cancer cell lines MIAPaCa-2 and PANC-1 for pancreatic drug testing, using the acid phosphatase assay. Drug efficacy testing showed that spheroids had much higher drug resistance than monolayers. This model, which is characteristically reproducible and easy and offers rapid handling, is the preferred choice for filling the gap between monolayer cell cultures and in vivo models in the process of drug development and testing for pancreatic cancer.

  20. A spheroid-based 3-D culture model for pancreatic cancer drug testing, using the acid phosphatase assay

    Directory of Open Access Journals (Sweden)

    Z. Wen

    2013-08-01

    Full Text Available Current therapy for pancreatic cancer is multimodal, involving surgery and chemotherapy. However, development of pancreatic cancer therapies requires a thorough evaluation of drug efficacy in vitro before animal testing and subsequent clinical trials. Compared to two-dimensional culture of cell monolayer, three-dimensional (3-D models more closely mimic native tissues, since the tumor microenvironment established in 3-D models often plays a significant role in cancer progression and cellular responses to the drugs. Accumulating evidence has highlighted the benefits of 3-D in vitro models of various cancers. In the present study, we have developed a spheroid-based, 3-D culture of pancreatic cancer cell lines MIAPaCa-2 and PANC-1 for pancreatic drug testing, using the acid phosphatase assay. Drug efficacy testing showed that spheroids had much higher drug resistance than monolayers. This model, which is characteristically reproducible and easy and offers rapid handling, is the preferred choice for filling the gap between monolayer cell cultures and in vivo models in the process of drug development and testing for pancreatic cancer.

  1. MAIN FEATURES OF A 3D GIS FOR A MONUMENTAL COMPLEX WITH AN HISTORICAL-CULTURAL RELEVANCE

    Directory of Open Access Journals (Sweden)

    A. Scianna

    2017-05-01

    Full Text Available The last achievements of technologies in geomatics especially in survey and restitution of 3D models (UAV/drones and laser scanner technologies generated new procedures and higher standards of quality in representation of archaeological sites. Together with Geomatics, the recent development of Information and Communication Technologies (ICT strongly contribute to document and the Cultural Heritage (CH. The representation and documentation of CH using these new technologies has became necessary in order to satisfy different needs: – for restorers in order to acquire a deep knowledge of the cultural good and to define possible strategies of restoration; – for the conservation of information, allowing to preserve the 3D geometry of the monumental complex with the integration of descriptions about architectural elements; – for touristic aims, giving the opportunity of sharing CH information on web, allowing users to visit and explore, in a virtual way, monumental complexes, acquiring information details about architectural elements or the history of monumental complex. Looking through these new scenarios, the development of a 3D Geographic Information System (GIS applied to a cultural good could be, today, an added value of fundamental importance for full description and data management of monumental complexes. In this work, the main features necessary for the correct construction of a 3D GIS of a monumental complex will be analyzed, with a particular focus on the possibilities for creating a standardized procedure to follow.

  2. 3D Visualization of Cultural Heritage Artefacts with Virtual Reality devices

    Science.gov (United States)

    Gonizzi Barsanti, S.; Caruso, G.; Micoli, L. L.; Covarrubias Rodriguez, M.; Guidi, G.

    2015-08-01

    Although 3D models are useful to preserve the information about historical artefacts, the potential of these digital contents are not fully accomplished until they are not used to interactively communicate their significance to non-specialists. Starting from this consideration, a new way to provide museum visitors with more information was investigated. The research is aimed at valorising and making more accessible the Egyptian funeral objects exhibited in the Sforza Castle in Milan. The results of the research will be used for the renewal of the current exhibition, at the Archaeological Museum in Milan, by making it more attractive. A 3D virtual interactive scenario regarding the "path of the dead", an important ritual in ancient Egypt, was realized to augment the experience and the comprehension of the public through interactivity. Four important artefacts were considered for this scope: two ushabty, a wooden sarcophagus and a heart scarab. The scenario was realized by integrating low-cost Virtual Reality technologies, as the Oculus Rift DK2 and the Leap Motion controller, and implementing a specific software by using Unity. The 3D models were implemented by adding responsive points of interest in relation to important symbols or features of the artefact. This allows highlighting single parts of the artefact in order to better identify the hieroglyphs and provide their translation. The paper describes the process for optimizing the 3D models, the implementation of the interactive scenario and the results of some test that have been carried out in the lab.

  3. Weak lensing reconstructions in 2D & 3D: implications for cluster studies

    CERN Document Server

    Leonard, Adrienne; Starck, Jean-Luc

    2015-01-01

    We compare the efficiency with which 2D and 3D weak lensing mass mapping techniques are able to detect clusters of galaxies using two state-of-the-art mass reconstruction techniques: MRLens in 2D and GLIMPSE in 3D. We simulate otherwise-empty cluster fields for 96 different virial mass-redshift combinations spanning the ranges $3\\times10^{13}h^{-1}M_\\odot \\le M_{vir}\\le 10^{15}h^{-1}M_\\odot$ and $0.05 \\le z_{\\rm cl} \\le 0.75$, and for each generate 1000 realisations of noisy shear data in 2D and 3D. For each field, we then compute the cluster (false) detection rate as the mean number of cluster (false) detections per reconstruction over the sample of 1000 reconstructions. We show that both MRLens and GLIMPSE are effective tools for the detection of clusters from weak lensing measurements, and provide comparable quality reconstructions at low redshift. At high redshift, GLIMPSE reconstructions offer increased sensitivity in the detection of clusters, yielding cluster detection rates up to a factor of $\\sim 10\\...

  4. Development of a 3D Tissue Culture-Based High-Content Screening Platform That Uses Phenotypic Profiling to Discriminate Selective Inhibitors of Receptor Tyrosine Kinases.

    NARCIS (Netherlands)

    Booij, T.H.; Klop, M.J.; Yan, K.; Szántai-Kis, C.; Szokol, B.; Orfi, L .; Water, van de B.; Keri, G.; Price, L.S.

    2016-01-01

    3D tissue cultures provide a more physiologically relevant context for the screening of compounds, compared with 2D cell cultures. Cells cultured in 3D hydrogels also show complex phenotypes, increasing the scope for phenotypic profiling. Here we describe a high-content screening platform that uses

  5. Development of a 3D Tissue Culture-Based High-Content Screening Platform That Uses Phenotypic Profiling to Discriminate Selective Inhibitors of Receptor Tyrosine Kinases.

    NARCIS (Netherlands)

    Booij, T.H.; Klop, M.J.; Yan, K.; Szántai-Kis, C.; Szokol, B.; Orfi, L .; Water, van de B.; Keri, G.; Price, L.S.

    2016-01-01

    3D tissue cultures provide a more physiologically relevant context for the screening of compounds, compared with 2D cell cultures. Cells cultured in 3D hydrogels also show complex phenotypes, increasing the scope for phenotypic profiling. Here we describe a high-content screening platform that uses

  6. Induced tauopathy in a novel 3D-culture model mediates neurodegenerative processes: a real-time study on biochips.

    Science.gov (United States)

    Seidel, Diana; Krinke, Dana; Jahnke, Heinz-Georg; Hirche, Anika; Kloß, Daniel; Mack, Till G A; Striggow, Frank; Robitzki, Andrea

    2012-01-01

    Tauopathies including Alzheimer's disease represent one of the major health problems of aging population worldwide. Therefore, a better understanding of tau-dependent pathologies and consequently, tau-related intervention strategies is highly demanded. In recent years, several tau-focused therapies have been proposed with the aim to stop disease progression. However, to develop efficient active pharmaceutical ingredients for the broad treatment of Alzheimer's disease patients, further improvements are necessary for understanding the detailed neurodegenerative processes as well as the mechanism and side effects of potential active pharmaceutical ingredients (API) in the neuronal system. In this context, there is a lack of suitable complex in vitro cell culture models recapitulating major aspects of taupathological degenerative processes in sufficient time and reproducible manner.Herewith, we describe a novel 3D SH-SY5Y cell-based, tauopathy model that shows advanced characteristics of matured neurons in comparison to monolayer cultures without the need of artificial differentiation promoting agents. Moreover, the recombinant expression of a novel highly pathologic fourfold mutated human tau variant lead to a fast and emphasized degeneration of neuritic processes. The neurodegenerative effects could be analyzed in real time and with high sensitivity using our unique microcavity array-based impedance spectroscopy measurement system. We were able to quantify a time- and concentration-dependent relative impedance decrease when Alzheimer's disease-like tau pathology was induced in the neuronal 3D cell culture model. In combination with the collected optical information, the degenerative processes within each 3D-culture could be monitored and analyzed. More strikingly, tau-specific regenerative effects caused by tau-focused active pharmaceutical ingredients could be quantitatively monitored by impedance spectroscopy.Bringing together our novel complex 3D cell culture

  7. Development of thermoresponsive non-woven 3D scaffold for smart cell culture

    CSIR Research Space (South Africa)

    Mahlangu, T

    2012-10-01

    Full Text Available .kashan.co.za] INTRODUCTION Conventional cell culture of adherent cells, based on the use of tissue culture polystyrene trays is an inefficient method to culture cells. The method employs the use of 2D surfaces and enzymatic treatment to release propagated cells...

  8. High-accuracy 3-D modeling of cultural heritage: the digitizing of Donatello's "Maddalena".

    Science.gov (United States)

    Guidi, Gabriele; Beraldin, J Angelo; Atzeni, Carlo

    2004-03-01

    Three-dimensional digital modeling of Heritage works of art through optical scanners, has been demonstrated in recent years with results of exceptional interest. However, the routine application of three-dimensional (3-D) modeling to Heritage conservation still requires the systematic investigation of a number of technical problems. In this paper, the acquisition process of the 3-D digital model of the Maddalena by Donatello, a wooden statue representing one of the major masterpieces of the Italian Renaissance which was swept away by the Florence flood of 1966 and successively restored, is described. The paper reports all the steps of the acquisition procedure, from the project planning to the solution of the various problems due to range camera calibration and to material non optically cooperative. Since the scientific focus is centered on the 3-D model overall dimensional accuracy, a methodology for its quality control is described. Such control has demonstrated how, in some situations, the ICP-based alignment can lead to incorrect results. To circumvent this difficulty we propose an alignment technique based on the fusion of ICP with close-range digital photogrammetry and a non-invasive procedure in order to generate a final accurate model. In the end detailed results are presented, demonstrating the improvement of the final model, and how the proposed sensor fusion ensure a pre-specified level of accuracy.

  9. A multifunctional 3D co-culture system for studies of mammary tissue morphogenesis and stem cell biology.

    Science.gov (United States)

    Campbell, Jonathan J; Davidenko, Natalia; Caffarel, Maria M; Cameron, Ruth E; Watson, Christine J

    2011-01-01

    Studies on the stem cell niche and the efficacy of cancer therapeutics require complex multicellular structures and interactions between different cell types and extracellular matrix (ECM) in three dimensional (3D) space. We have engineered a 3D in vitro model of mammary gland that encompasses a defined, porous collagen/hyaluronic acid (HA) scaffold forming a physiologically relevant foundation for epithelial and adipocyte co-culture. Polarized ductal and acinar structures form within this scaffold recapitulating normal tissue morphology in the absence of reconstituted basement membrane (rBM) hydrogel. Furthermore, organoid developmental outcome can be controlled by the ratio of collagen to HA, with a higher HA concentration favouring acinar morphological development. Importantly, this culture system recapitulates the stem cell niche as primary mammary stem cells form complex organoids, emphasising the utility of this approach for developmental and tumorigenic studies using genetically altered animals or human biopsy material, and for screening cancer therapeutics for personalised medicine.

  10. A multifunctional 3D co-culture system for studies of mammary tissue morphogenesis and stem cell biology.

    Directory of Open Access Journals (Sweden)

    Jonathan J Campbell

    Full Text Available Studies on the stem cell niche and the efficacy of cancer therapeutics require complex multicellular structures and interactions between different cell types and extracellular matrix (ECM in three dimensional (3D space. We have engineered a 3D in vitro model of mammary gland that encompasses a defined, porous collagen/hyaluronic acid (HA scaffold forming a physiologically relevant foundation for epithelial and adipocyte co-culture. Polarized ductal and acinar structures form within this scaffold recapitulating normal tissue morphology in the absence of reconstituted basement membrane (rBM hydrogel. Furthermore, organoid developmental outcome can be controlled by the ratio of collagen to HA, with a higher HA concentration favouring acinar morphological development. Importantly, this culture system recapitulates the stem cell niche as primary mammary stem cells form complex organoids, emphasising the utility of this approach for developmental and tumorigenic studies using genetically altered animals or human biopsy material, and for screening cancer therapeutics for personalised medicine.

  11. State-of-the-art of 3D cultures (organs-on-a-chip) in safety testing and pathophysiology

    OpenAIRE

    Alépée, N.; Bahinski, A.; Daneshian, M.; B. de Wever; Fritsche, E.; Goldberg, A.; Hansmann, J.; Hartung, T; Haycock, J; Hogberg, H.; Hoelting, L.; Kelm, J M; Kadereit, S.; McVey, E.; Landsiedel, R.

    2014-01-01

    Integrated approaches using different in vitro methods in combination with bioinformatics can (i) increase the success rate and speed of drug development; (ii) improve the accuracy of toxicological risk assessment; and (iii) increase our understanding of disease. An important building block of this strategy that has emerged during the last years are threedimensional (3D) cell culture models. The majority of these models are organotypic, i.e., they aim to reproduce major functions of an organ ...

  12. Integrating 3D seismic curvature and curvature gradient attributes for fracture characterization: Methodologies and interpretational implications

    Energy Technology Data Exchange (ETDEWEB)

    Gao, Dengliang

    2013-03-01

    In 3D seismic interpretation, curvature is a popular attribute that depicts the geometry of seismic reflectors and has been widely used to detect faults in the subsurface; however, it provides only part of the solutions to subsurface structure analysis. This study extends the curvature algorithm to a new curvature gradient algorithm, and integrates both algorithms for fracture detection using a 3D seismic test data set over Teapot Dome (Wyoming). In fractured reservoirs at Teapot Dome known to be formed by tectonic folding and faulting, curvature helps define the crestal portion of the reservoirs that is associated with strong seismic amplitude and high oil productivity. In contrast, curvature gradient helps better define the regional northwest-trending and the cross-regional northeast-trending lineaments that are associated with weak seismic amplitude and low oil productivity. In concert with previous reports from image logs, cores, and outcrops, the current study based on an integrated seismic curvature and curvature gradient analysis suggests that curvature might help define areas of enhanced potential to form tensile fractures, whereas curvature gradient might help define zones of enhanced potential to develop shear fractures. In certain fractured reservoirs such as at Teapot Dome where faulting and fault-related folding contribute dominantly to the formation and evolution of fractures, curvature and curvature gradient attributes can be potentially applied to differentiate fracture mode, to predict fracture intensity and orientation, to detect fracture volume and connectivity, and to model fracture networks.

  13. Hydrogel Based 3-Dimensional (3D System for Toxicity and High-Throughput (HTP Analysis for Cultured Murine Ovarian Follicles.

    Directory of Open Access Journals (Sweden)

    Hong Zhou

    Full Text Available Various toxicants, drugs and their metabolites carry potential ovarian toxicity. Ovarian follicles, the functional unit of the ovary, are susceptible to this type of damage at all stages of their development. However, despite of the large scale of potential negative impacts, assays that study ovarian toxicity are limited. Exposure of cultured ovarian follicles to toxicants of interest served as an important tool for evaluation of toxic effects for decades. Mouse follicles cultured on the bottom of a culture dish continue to serve an important approach for mechanistic studies. In this paper, we demonstrated the usefulness of a hydrogel based 3-dimensional (3D mouse ovarian follicle culture as a tool to study ovarian toxicity in a different setup. The 3D in vitro culture, based on fibrin alginate interpenetrating network (FA-IPN, preserves the architecture of the ovarian follicle and physiological structure-function relationship. We applied the novel 3D high-throughput (HTP in vitro ovarian follicle culture system to study the ovotoxic effects of an anti-cancer drug, Doxorobucin (DXR. The fibrin component in the system is degraded by plasmin and appears as a clear circle around the encapsulated follicle. The degradation area of the follicle is strongly correlated with follicle survival and growth. To analyze fibrin degradation in a high throughput manner, we created a custom MATLAB® code that converts brightfield micrographs of follicles encapsulated in FA-IPN to binary images, followed by image analysis. We did not observe any significant difference between manually processed images to the automated MATLAB® method, thereby confirming that the automated program is suitable to measure fibrin degradation to evaluate follicle health. The cultured follicles were treated with DXR at concentrations ranging from 0.005 nM to 200 nM, corresponding to the therapeutic plasma levels of DXR in patients. Follicles treated with DXR demonstrated decreased

  14. Hydrogel Based 3-Dimensional (3D) System for Toxicity and High-Throughput (HTP) Analysis for Cultured Murine Ovarian Follicles.

    Science.gov (United States)

    Zhou, Hong; Malik, Malika Amattullah; Arab, Aarthi; Hill, Matthew Thomas; Shikanov, Ariella

    2015-01-01

    Various toxicants, drugs and their metabolites carry potential ovarian toxicity. Ovarian follicles, the functional unit of the ovary, are susceptible to this type of damage at all stages of their development. However, despite of the large scale of potential negative impacts, assays that study ovarian toxicity are limited. Exposure of cultured ovarian follicles to toxicants of interest served as an important tool for evaluation of toxic effects for decades. Mouse follicles cultured on the bottom of a culture dish continue to serve an important approach for mechanistic studies. In this paper, we demonstrated the usefulness of a hydrogel based 3-dimensional (3D) mouse ovarian follicle culture as a tool to study ovarian toxicity in a different setup. The 3D in vitro culture, based on fibrin alginate interpenetrating network (FA-IPN), preserves the architecture of the ovarian follicle and physiological structure-function relationship. We applied the novel 3D high-throughput (HTP) in vitro ovarian follicle culture system to study the ovotoxic effects of an anti-cancer drug, Doxorobucin (DXR). The fibrin component in the system is degraded by plasmin and appears as a clear circle around the encapsulated follicle. The degradation area of the follicle is strongly correlated with follicle survival and growth. To analyze fibrin degradation in a high throughput manner, we created a custom MATLAB® code that converts brightfield micrographs of follicles encapsulated in FA-IPN to binary images, followed by image analysis. We did not observe any significant difference between manually processed images to the automated MATLAB® method, thereby confirming that the automated program is suitable to measure fibrin degradation to evaluate follicle health. The cultured follicles were treated with DXR at concentrations ranging from 0.005 nM to 200 nM, corresponding to the therapeutic plasma levels of DXR in patients. Follicles treated with DXR demonstrated decreased survival rate in

  15. Digital Inventory and Documentation of Korea's Important Cultural Properties Using 3D Laser Scanning

    Science.gov (United States)

    Dongseok, K.; Gyesoo, K.; Siro, K.; Eunhwa, K.

    2015-08-01

    As a country with 11 properties included on the World Heritage List and approximately 12,000 important cultural properties, Korea has been continuously carrying out the inventory and documentation of cultural properties to conserve and manage them since the 1960s. The inventory of cultural properties had been carried out by making and managing a register which recorded basic information mainly on state-designated cultural properties such as their size, quantity, and location. The documentation of cultural properties was also carried out by making measured drawings. However, the inventory and documentation done under the previous analog method had a limit to the information it could provide for the effective conservation and management of cultural properties. Moreover, in recent times important cultural properties have frequently been damaged by man-made and natural disasters such as arson, forest fires, and floods, so an alternative was required. Accordingly, Korea actively introduced digital techniques led by the government for the inventory and documentation of important cultural properties. In this process, the government established the concept of a digital set, built a more efficie nt integrated data management system, and created standardized guidelines to maximize the effectiveness of data acquisition, management, and utilization that greatly increased the level of digital inventory, documentation, and archiving.

  16. Researching Interplay between 3D-Materials and Young Children in Socio-Cultural Contexts

    Directory of Open Access Journals (Sweden)

    Biljana C. Fredriksen

    2011-11-01

    Full Text Available This text presents empirical and interpretative methodological inquiry in a study of young children’s interplay with three-dimensional (3D materials in early childhood educational settings. Recent acknowledgement of young children as competent individuals challenges forms of research with them. Respecting their views and competence demands that they are treated as actors and not objects of research. At the same time, young children are vulnerable and need to be protected from harm, for example when they use tools. This combination of children’s competence and vulnerability challenges research ethics and methods. This article discusses the following question: How to conduct ethical and valid research in sloyd education with young children?The study was carried out in a Norwegian Early Childhood Education Centre, ECEC, using a multiple case study approach. To be able to understand young children’s experiences, the researcher positioned herself inside the educational contexts taking the role of an A/R/T-ographer. Ten case contexts were conducted, in which pairs of different children played with 3D-materials. The cases were filmed and the video-material was analyzed both contextually and in a cross-case manner.Keywords: method, sloyd, early childhood education, video observation, three-dimensional materialsURN:NBN:no-29956

  17. Network dynamics of 3D engineered neuronal cultures: a new experimental model for in-vitro electrophysiology.

    Science.gov (United States)

    Frega, Monica; Tedesco, Mariateresa; Massobrio, Paolo; Pesce, Mattia; Martinoia, Sergio

    2014-06-30

    Despite the extensive use of in-vitro models for neuroscientific investigations and notwithstanding the growing field of network electrophysiology, all studies on cultured cells devoted to elucidate neurophysiological mechanisms and computational properties, are based on 2D neuronal networks. These networks are usually grown onto specific rigid substrates (also with embedded electrodes) and lack of most of the constituents of the in-vivo like environment: cell morphology, cell-to-cell interaction and neuritic outgrowth in all directions. Cells in a brain region develop in a 3D space and interact with a complex multi-cellular environment and extracellular matrix. Under this perspective, 3D networks coupled to micro-transducer arrays, represent a new and powerful in-vitro model capable of better emulating in-vivo physiology. In this work, we present a new experimental paradigm constituted by 3D hippocampal networks coupled to Micro-Electrode-Arrays (MEAs) and we show how the features of the recorded network dynamics differ from the corresponding 2D network model. Further development of the proposed 3D in-vitro model by adding embedded functionalized scaffolds might open new prospects for manipulating, stimulating and recording the neuronal activity to elucidate neurophysiological mechanisms and to design bio-hybrid microsystems.

  18. Advances in 3D cell culture technologies enabling tissue-like structures to be created in vitro.

    Science.gov (United States)

    Knight, Eleanor; Przyborski, Stefan

    2015-12-01

    Research in mammalian cell biology often relies on developing in vitro models to enable the growth of cells in the laboratory to investigate a specific biological mechanism or process under different test conditions. The quality of such models and how they represent the behavior of cells in real tissues plays a critical role in the value of the data produced and how it is used. It is particularly important to recognize how the structure of a cell influences its function and how co-culture models can be used to more closely represent the structure of real tissue. In recent years, technologies have been developed to enhance the way in which researchers can grow cells and more readily create tissue-like structures. Here we identify the limitations of culturing mammalian cells by conventional methods on two-dimensional (2D) substrates and review the popular approaches currently available that enable the development of three-dimensional (3D) tissue models in vitro. There are now many ways in which the growth environment for cultured cells can be altered to encourage 3D cell growth. Approaches to 3D culture can be broadly categorized into scaffold-free or scaffold-based culture systems, with scaffolds made from either natural or synthetic materials. There is no one particular solution that currently satisfies all requirements and researchers must select the appropriate method in line with their needs. Using such technology in conjunction with other modern resources in cell biology (e.g. human stem cells) will provide new opportunities to create robust human tissue mimetics for use in basic research and drug discovery. Application of such models will contribute to advancing basic research, increasing the predictive accuracy of compounds, and reducing animal usage in biomedical science.

  19. Collagen esterification enhances the function and survival of pancreatic β cells in 2D and 3D culture systems

    Energy Technology Data Exchange (ETDEWEB)

    Ko, Jae Hyung [Regenerative Medicine Research Center, Dalim Tissen Co., LTD., 383-93, Yonnam-Dong, Mapo-gu, Seoul (Korea, Republic of); Kim, Yang Hee [Regenerative Medicine Research Center, Dalim Tissen Co., LTD., 383-93, Yonnam-Dong, Mapo-gu, Seoul (Korea, Republic of); Asan Institute for Life Science, 388-1 Pungnap-2 Dong, Songpa-gu, Seoul (Korea, Republic of); Jeong, Seong Hee; Lee, Song [Asan Institute for Life Science, 388-1 Pungnap-2 Dong, Songpa-gu, Seoul (Korea, Republic of); Park, Si-Nae [Regenerative Medicine Research Center, Dalim Tissen Co., LTD., 383-93, Yonnam-Dong, Mapo-gu, Seoul (Korea, Republic of); Shim, In Kyong, E-mail: shimiink@gmail.com [Asan Institute for Life Science, 388-1 Pungnap-2 Dong, Songpa-gu, Seoul (Korea, Republic of); Kim, Song Cheol, E-mail: drksc@amc.seoul.kr [Asan Institute for Life Science, 388-1 Pungnap-2 Dong, Songpa-gu, Seoul (Korea, Republic of); Department of Surgery, University of Ulsan College of Medicine & Asan Medical Center, 388-1 Pungnap-2 Dong, Songpa-gu, Seoul (Korea, Republic of)

    2015-08-07

    Collagen, one of the most important components of the extracellular matrix (ECM), may play a role in the survival of pancreatic islet cells. In addition, chemical modifications that change the collagen charge profile to a net positive charge by esterification have been shown to increase the adhesion and proliferation of various cell types. The purpose of this study was to characterize and compare the effects of native collagen (NC) and esterified collagen (EC) on β cell function and survival. After isolation by the collagenase digestion technique, rat islets were cultured with NC and EC in 2 dimensional (2D) and 3 dimensional (3D) environments for a long-term duration in vitro. The cells were assessed for islet adhesion, morphology, viability, glucose-induced insulin secretion, and mRNA expression of glucose metabolism-related genes, and visualized by scanning electron microscopy (SEM). Islet cells attached tightly in the NC group, but islet cell viability was similar in both the NC and EC groups. Glucose-stimulated insulin secretion was higher in the EC group than in the NC group in both 2D and 3D culture. Furthermore, the mRNA expression levels of glucokinase in the EC group were higher than those in the NC group and were associated with glucose metabolism and insulin secretion. Finally, SEM observation confirmed that islets had more intact component cells on EC sponges than on NC sponges. These results indicate that modification of collagen may offer opportunities to improve function and viability of islet cells. - Highlights: • We changed the collagen charge profile to a net positive charge by esterification. • Islets cultured on esterified collagen improved survival in both 2D and 3D culture. • Islets cultured on esterified collagen enhanced glucose-stimulated insulin release. • High levels of glucokinase mRNA may be associated with increased insulin release.

  20. Development of thermoresponsive poly(propylene-g-N-isopropylacrylamide) non-woven 3D scaffold for smart cell culture using oxyfluorination-assisted graft polymerisation

    CSIR Research Space (South Africa)

    Chetty, AS

    2013-02-01

    Full Text Available Growing cells on 3D scaffolds is far superior to the conventional 2D monolayer culture method. In this study, a novel 3D thermoresponsive poly(propylene-g-N-isopropylacrylamide) (PP-g-PNIPAAm) nonwoven fabric (gNWF) was developed for cell culture...

  1. Recording, Visualization and Documentation of 3D Spatial Data for Monitoring Topography in Areas of Cultural Heritage

    Science.gov (United States)

    Maravelakis, Emmanouel; Konstantaras, Antonios; Axaridou, Anastasia; Chrysakis, Ioannis; Xinogalos, Michalis

    2014-05-01

    This research investigates the application of new system for 3D documentation of land degradation and its effect [1,2] on areas of cultural heritage via complete 3D data acquisition, 3D modeling and metadata recording using terrestrial laser scanners (TLS) [3,4,5]. As land degradation progresses through time it is important to be able to map and exactly replicate with great precision the entire 3D shape of the physical objects of interest, such as landslides, ground erosion, river boundaries, mad accumulation, etc. [1,2] TLS enables the extraction and recording of a very large number of points in space with great precision and without the need for any physical contact with the object of interest. Field specialists can then examine the produced models and comment on them both on the overall object of interest and on specific features of it by inserting annotations on certain parts of the model [6]. This process could be proven to be very cost effective as it can be repeated as often as necessary and produce a well catalogued documentation of the progress of land degradation at particular areas. The problem with repeating TLS models lies on the various types of hardware equipment and software systems that might be used for the extraction of point clouds, and the different people that might be called to analyze the findings. These often result in a large volume of interim and final products with little if no standardization, multiple different metadata and vague documentation [7], which makes metadata recordings [8] crucial both for one scientist to be able to follow upon the work of the other as well as being able to repeat the same work when deemed necessary. This makes the need for a repository tool proposed by the authors essential in order to record all work that is done in every TLS scanning, and makes the technology accessible to scientists of various different fields [9,10], eg. geologists, physicists, topographers, remote sensing engineers, archaeologists etc

  2. Differentiation of neuroepithelial stem cells into functional dopaminergic neurons in 3D microfluidic cell culture.

    Science.gov (United States)

    Moreno, Edinson Lucumi; Hachi, Siham; Hemmer, Kathrin; Trietsch, Sebastiaan J; Baumuratov, Aidos S; Hankemeier, Thomas; Vulto, Paul; Schwamborn, Jens C; Fleming, Ronan M T

    2015-06-07

    A hallmark of Parkinson's disease is the progressive loss of nigrostriatal dopaminergic neurons. We derived human neuroepithelial cells from induced pluripotent stem cells and successfully differentiated them into dopaminergic neurons within phase-guided, three-dimensional microfluidic cell culture bioreactors. After 30 days of differentiation within the microfluidic bioreactors, in situ morphological, immunocytochemical and calcium imaging confirmed the presence of dopaminergic neurons that were spontaneously electrophysiologically active, a characteristic feature of nigrostriatal dopaminergic neurons in vivo. Differentiation was as efficient as in macroscopic culture, with up to 19% of differentiated neurons immunoreactive for tyrosine hydroxylase, the penultimate enzyme in the synthesis of dopamine. This new microfluidic cell culture model integrates the latest innovations in developmental biology and microfluidic cell culture to generate a biologically realistic and economically efficient route to personalised drug discovery for Parkinson's disease.

  3. Real-Time Cell Cycle Imaging in a 3D Cell Culture Model of Melanoma.

    Science.gov (United States)

    Spoerri, Loredana; Beaumont, Kimberley A; Anfosso, Andrea; Haass, Nikolas K

    2017-01-01

    Aberrant cell cycle progression is a hallmark of solid tumors; therefore, cell cycle analysis is an invaluable technique to study cancer cell biology. However, cell cycle progression has been most commonly assessed by methods that are limited to temporal snapshots or that lack spatial information. Here, we describe a technique that allows spatiotemporal real-time tracking of cell cycle progression of individual cells in a multicellular context. The power of this system lies in the use of 3D melanoma spheroids generated from melanoma cells engineered with the fluorescent ubiquitination-based cell cycle indicator (FUCCI). This technique allows us to gain further and more detailed insight into several relevant aspects of solid cancer cell biology, such as tumor growth, proliferation, invasion, and drug sensitivity.

  4. An Interdisciplinary Conservation Module for Condition Survey on Cultural Heritages with a 3d Information System

    Science.gov (United States)

    Pedelì, C.

    2013-07-01

    In order to make the most of the digital outsourced documents, based on new technologies (e.g.: 3D LASER scanners, photogrammetry, etc.), a new approach was followed and a new ad hoc information system was implemented. The obtained product allow to the final user to reuse and manage the digital documents providing graphic tools and an integrated specific database to manage the entire documentation and conservation process, starting from the condition assessment until the conservation / restoration work. The system is organised on two main modules: Archaeology and Conservation. This paper focus on the features and the advantages of the second one. In particular it is emphasized its logical organisation, the possibility to easily mapping by using a very precise 3D metric platform, to benefit of the integrated relational database which allows to well organise, compare, keep and manage different kind of information at different level. Conservation module can manage along the time the conservation process of a site, monuments, object or excavation and conservation work in progress. An alternative approach called OVO by the author of this paper, force the surveyor to observe and describe the entity decomposing it on functional components, materials and construction techniques. Some integrated tools as the "ICOMOS-ISCS Illustrated glossary … " help the user to describe pathologies with a unified approach and terminology. Also the conservation project phase is strongly supported to envision future intervention and cost. A final section is devoted to record the conservation/restoration work already done or in progress. All information areas of the conservation module are interconnected to each other to allows to the system a complete interchange of graphic and alphanumeric data. The conservation module it self is connected to the archaeological one to create an interdisciplinary daily tool.

  5. High-Throughput Cancer Cell Sphere Formation for Characterizing the Efficacy of Photo Dynamic Therapy in 3D Cell Cultures

    Science.gov (United States)

    Chen, Yu-Chih; Lou, Xia; Zhang, Zhixiong; Ingram, Patrick; Yoon, Euisik

    2015-07-01

    Photodynamic therapy (PDT), wherein light sensitive non-toxic agents are locally and selectively activated using light, has emerged as an appealing alternative to traditional cancer chemotherapy. Yet to date, PDT efficacy has been mostly characterized using 2D cultures. Compared to 2D cultures, 3D sphere culture generates unique spatial distributions of nutrients and oxygen for the cells that better mimics the in-vivo conditions. Using a novel polyHEMA (non-adherent polymer) fabrication process, we developed a microfluidic sphere formation platform that can (1) generate 1,024 uniform (size variation characterized the different responses in 2D and 3D cell culture to PDT. Furthermore, we investigated the treatment resistance effect in cancer cells induced by tumor associated fibroblasts (CAF). Although the CAFs can enhance the resistance to traditional chemotherapy agents, no significant difference in PDT was observed. The preliminary results suggest that the PDT can be an attractive alternative cancer therapy, which is less affected by the therapeutic resistance induced by cancer associated cells.

  6. Protein-engineered scaffolds for in vitro 3D culture of primary adult intestinal organoids.

    Science.gov (United States)

    DiMarco, Rebecca L; Dewi, Ruby E; Bernal, Gabriela; Kuo, Calvin; Heilshorn, Sarah C

    2015-10-15

    Though in vitro culture of primary intestinal organoids has gained significant momentum in recent years, little has been done to investigate the impact of microenvironmental cues provided by the encapsulating matrix on the growth and development of these fragile cultures. In this work, the impact of various in vitro culture parameters on primary adult murine organoid formation and growth are analyzed with a focus on matrix properties and geometric culture configuration. The air-liquid interface culture configuration was found to result in enhanced organoid formation relative to a traditional submerged configuration. Additionally, through use of a recombinantly engineered extracellular matrix (eECM), the effects of biochemical and biomechanical cues were independently studied. Decreasing mechanical stiffness and increasing cell adhesivity were found to increase organoid yield. Tuning of eECM properties was used to obtain organoid formation efficiency values identical to those observed in naturally harvested collagen I matrices but within a stiffer construct with improved ease of physical manipulation. Increased ability to remodel the surrounding matrix through mechanical or enzymatic means was also shown to enhance organoid formation. As the engineering and tunability of recombinant matrices is essentially limitless, continued property optimization may result in further improved matrix performance and may help to identify additional microenvironmental cues that directly impact organoid formation, development, differentiation, and functional behavior. Continued culture of primary organoids in recombinant matrices could therefore prove to be largely advantageous in the field of intestinal tissue engineering for applications in regenerative medicine and in vitro tissue mimics.

  7. Titan's past and future: 3D modeling of a pure nitrogen atmosphere and geological implications

    CERN Document Server

    Charnay, Benjamin; Tobie, Gabriel; Sotin, Christophe; Wordsworth, Robin

    2014-01-01

    Several clues indicate that Titan's atmosphere has been depleted in methane during some period of its history, possibly as recently as 0.5-1 billion years ago. It could also happen in the future. Under these conditions, the atmosphere becomes only composed of nitrogen with a range of temperature and pressure allowing liquid or solid nitrogen to condense. Here, we explore these exotic climates throughout Titan's history with a 3D Global Climate Model (GCM) including the nitrogen cycle and the radiative effect of nitrogen clouds. We show that for the last billion years, only small polar nitrogen lakes should have formed. Yet, before 1 Ga, a significant part of the atmosphere could have condensed, forming deep nitrogen polar seas, which could have flowed and flooded the equatorial regions. Alternatively, nitrogen could be frozen on the surface like on Triton, but this would require an initial surface albedo higher than 0.65 at 4 Ga. Such a state could be stable even today if nitrogen ice albedo is higher than th...

  8. 3D modeling of GJ1214b's atmosphere: formation of inhomogeneous high clouds and observational implications

    CERN Document Server

    Charnay, Benjamin; Misra, Amit; Leconte, Jérémy; Arney, Giada

    2015-01-01

    The warm sub-Neptune GJ1214b has a featureless transit spectrum which may be due to the presence of high and thick clouds or haze. Here, we simulate the atmosphere of GJ1214b with a 3D General Circulation Model for cloudy hydrogen-dominated atmospheres, including cloud radiative effects. We show that the atmospheric circulation is strong enough to transport micrometric cloud particles to the upper atmosphere and generally leads to a minimum of cloud at the equator. By scattering stellar light, clouds increase the planetary albedo to 0.4-0.6 and cool the atmosphere below 1 mbar. However, the heating by ZnS clouds leads to the formation of a stratospheric thermal inversion above 10 mbar, with temperatures potentially high enough on the dayside to evaporate KCl clouds. We show that flat transit spectra consistent with HST observations are possible if cloud particle radii are around 0.5 micron, and that such clouds should be optically thin at wavelengths > 3 microns. Using simulated cloudy atmospheres that fit th...

  9. 3D Body Wave Velocity Tomography in Southern Peru: Seismotectonic Implications

    Science.gov (United States)

    Gallego, A.; Perez, J.; David, C.; Comte, D.; Charrier, R.; Dorbath, L.

    2004-12-01

    The studied region corresponds to the southern segment of the 1868 rupture area, that did not break with the last Mw=8.4 Arequipa earthquake in southern Peru. A temporary network of 19 short period, continuous recording seismic stations was deployed (16.5° -18.5° S; 69.5° -72° W) between December 2002-March 2003 in this region. The 1093 select events were used for a joint hypocentral and velocity structure inversion. The first obtained 1D velocity model was used as the initial model for the 3D inversion, consisting in 231 blocks distributed along layers separated by 10 km for depths lower than 80 km, and by 20 km for depths between 80 and 160 km. North of the Arica Bend, between the Coast and Andean Range the high Vp velocity and Vp/Vs ratio observed at depths lower than 10 km could represent the Precambrian basement uplifted by the Incapuquio sinistral fault system, which develops a positive flower structure typical for transpressional zones, that raised the basement. This behavior is in good agreement with the uplift of the Cambrian metamorphic complex, in the Precordillera south of the Arica Bend (northern Chile), by the west-vergent thrust system. Beneath the anomalous high velocities (between 20 and 30 km depth), a low velocity zone (Vpearthquake post-seismic period.

  10. Co-culture of 3D tumor spheroids with fibroblasts as a model for epithelial–mesenchymal transition in vitro

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    Kim, Sun-Ah, E-mail: j.sarah.k@gmail.com [Department of Biomedicine & Health Sciences, College of Medicine, The Catholic University of Korea, Seoul 137-701 (Korea, Republic of); Lee, Eun Kyung, E-mail: leeek@catholic.ac.kr [Department of Biochemistry, College of Medicine, The Catholic University of Korea, Seoul 137-701 (Korea, Republic of); Cancer Evolution Research Center, College of Medicine, The Catholic University of Korea, Seoul 137-701 (Korea, Republic of); Kuh, Hyo-Jeong, E-mail: hkuh@catholic.ac.kr [Department of Biomedicine & Health Sciences, College of Medicine, The Catholic University of Korea, Seoul 137-701 (Korea, Republic of); Cancer Evolution Research Center, College of Medicine, The Catholic University of Korea, Seoul 137-701 (Korea, Republic of)

    2015-07-15

    Epithelial–mesenchymal transition (EMT) acts as a facilitator of metastatic dissemination in the invasive margin of malignant tumors where active tumor–stromal crosstalks take place. Co-cultures of cancer cells with cancer-associated fibroblasts (CAFs) are often used as in vitro models of EMT. We established a tumor–fibroblast proximity co-culture using HT-29 tumor spheroids (TSs) with CCD-18co fibroblasts. When co-cultured with TSs, CCD-18co appeared activated, and proliferative activity as well as cell migration increased. Expression of fibronectin increased whereas laminin and type I collagen decreased in TSs co-cultured with fibroblasts compared to TSs alone, closely resembling the margin of in vivo xenograft tissue. Active TGFβ1 in culture media significantly increased in TS co-cultures but not in 2D co-cultures of cancer cells–fibroblasts, indicating that 3D context-associated factors from TSs may be crucial to crosstalks between cancer cells and fibroblasts. We also observed in TSs co-cultured with fibroblasts increased expression of α-SMA, EGFR and CTGF; reduced expression of membranous β-catenin and E-cadherin, together suggesting an EMT-like changes similar to a marginal region of xenograft tissue in vivo. Overall, our in vitro TS–fibroblast proximity co-culture mimics the EMT-state of the invasive margin of in vivo tumors in early metastasis. - Highlights: • An adjacent co-culture of tumor spheroids and fibroblasts is presented as EMT model. • Activation of fibroblasts and increased cell migration were shown in co-culture. • Expression of EMT-related factors in co-culture was similar to that in tumor tissue. • Crosstalk between spheroids and fibroblasts was demonstrated by secretome analysis.

  11. Between the Real and the Virtual: 3D visualization in the Cultural Heritage domain - expectations and prospects

    Directory of Open Access Journals (Sweden)

    Sorin Hermon

    2011-05-01

    Full Text Available The paper discusses two uses of 3D Visualization and Virtual Reality (hereafter VR of Cultural Heritage (CH assets: a less used one, in the archaeological / historical research and a more frequent one, as a communication medium in CH museums. While technological effort has been mainly invested in improving the “accuracy” of VR (determined as how truthfully it reproduces the “CH reality”, issues related to scientific requirements, (data transparency, separation between “real” and “virtual”, etc., are largely neglected, or at least not directly related to the 3D outcome, which may explain why, after more than twenty years of producing VR models, they are still rarely used in the archaeological research. The paper will present a proposal for developing VR tools as such as to be meaningful CH research tools as well as a methodology for designing VR outcomes to be used as a communication medium in CH museums.

  12. Web-based Visualization and Query of semantically segmented multiresolution 3D Models in the Field of Cultural Heritage

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    Auer, M.; Agugiaro, G.; Billen, N.; Loos, L.; Zipf, A.

    2014-05-01

    Many important Cultural Heritage sites have been studied over long periods of time by different means of technical equipment, methods and intentions by different researchers. This has led to huge amounts of heterogeneous "traditional" datasets and formats. The rising popularity of 3D models in the field of Cultural Heritage in recent years has brought additional data formats and makes it even more necessary to find solutions to manage, publish and study these data in an integrated way. The MayaArch3D project aims to realize such an integrative approach by establishing a web-based research platform bringing spatial and non-spatial databases together and providing visualization and analysis tools. Especially the 3D components of the platform use hierarchical segmentation concepts to structure the data and to perform queries on semantic entities. This paper presents a database schema to organize not only segmented models but also different Levels-of-Details and other representations of the same entity. It is further implemented in a spatial database which allows the storing of georeferenced 3D data. This enables organization and queries by semantic, geometric and spatial properties. As service for the delivery of the segmented models a standardization candidate of the OpenGeospatialConsortium (OGC), the Web3DService (W3DS) has been extended to cope with the new database schema and deliver a web friendly format for WebGL rendering. Finally a generic user interface is presented which uses the segments as navigation metaphor to browse and query the semantic segmentation levels and retrieve information from an external database of the German Archaeological Institute (DAI).

  13. Long-term immunologically competent human peripheral lymphoid tissue cultures in a 3D bioreactor

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    Kuzin, Igor; Sun, Hongliang; Moshkani, Safiekhatoon; Feng, Changyong; Mantalaris, Athanasios; Wu, JH David; Bottaro, Andrea

    2011-01-01

    Peripheral lymphoid organs (PLOs), the primary sites of development of adaptive immune responses, display a complex structural organization reflecting separation of cellular subsets (e.g. T and B lymphocytes) and functional compartments which is critical for immune function. The generation of in vitro culture systems capable of recapitulating salient features of PLOs for experimental, biotechnological and clinical applications would be highly desirable, but has been hampered so far by the complexity of these systems. We have previously developed a three-dimensional bioreactor system for long-term, functional culture of human bone marrow cells on macroporous microspheres in a packed-bed bioreactor with frequent medium change. Here we adapt the same system for culture of human primary cells from PLOs (tonsil) in the absence of specific exogenous growth factors or activators. Cells in this system displayed higher viability over several weeks, and maintain population diversity and cell surface markers largely comparable to primary cells. Light microscopy showed cells organizing in large diverse clusters within the scaffold pores and presence of B cell-enriched areas. Strikingly, these cultures generated a significant number of antibody-producing B cells when challenged with a panel of diverse antigens, as expected from a lymphoid tissue. Thus the three-dimensional tonsil bioreactor culture system may serve as a useful model of PLOs by recapitulating their structural organization and function ex vivo. PMID:21309085

  14. 3D Cell Culture in a Self-Assembled Nanofiber Environment

    Science.gov (United States)

    Gubbe, John D.; Brekke, John H.

    2016-01-01

    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. PMID:27632425

  15. 3D SURVEY AND AUGMENTED REALITY FOR CULTURAL HERITAGE. THE CASE STUDY OF AURELIAN WALL AT CASTRA PRAETORIA IN ROME

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    M. Canciani

    2016-06-01

    Full Text Available The development of close-range photogrammetry has produced a lot of new possibility to study cultural heritage. 3D data acquired with conventional and low cost cameras can be used to document, investigate the full appearance, materials and conservation status, to help the restoration process and identify intervention priorities. At the same time, with 3D survey a lot of three-dimensional data are collected and analyzed by researchers, but there are a very few possibility of 3D output. The augmented reality is one of this possible output with a very low cost technology but a very interesting result. Using simple mobile technology (for iPad and Android Tablets and shareware software (in the case presented “Augment” it is possible to share and visualize a large number of 3D models with your own device. The case study presented is a part of an architecture graduate thesis, made in Rome at Department of Architecture of Roma Tre University. We have developed a photogrammetric survey to study the Aurelian Wall at Castra Praetoria in Rome. The surveys of 8000 square meters of surface have allowed to identify stratigraphy and construction phases of a complex portion of Aurelian Wall, specially about the Northern door of Castra. During this study, the data coming out of 3D survey (photogrammetric and topographic, are stored and used to create a reverse 3D model, or virtual reconstruction, of the Northern door of Castra. This virtual reconstruction shows the door in the Tiberian period, nowadays it's totally hidden by a curtain wall but, little and significative architectural details allow to know its original feature. The 3D model of the ancient walls has been mapped with the exact type of bricks and mortar, oriented and scaled according to the existing one to use augmented reality. Finally, two kind of application have been developed, one on site, were you can see superimposed the virtual reconstruction on the existing walls using the image

  16. 3-D seismic tomography of the lithosphere and its geodynamic implications beneath the northeast India region

    Science.gov (United States)

    Raoof, J.; Mukhopadhyay, S.; Koulakov, I.; Kayal, J. R.

    2017-05-01

    We have evolved 3-D seismic velocity structures in northeast India region and its adjoining areas to understand the geodynamic processes of Indian lithosphere that gently underthrusts under the Himalayas and steeply subducts below the Indo-Burma Ranges. The region is tectonically buttressed between the Himalayan arc to the north and the Indo-Burmese arc to the east. The tomographic image shows heterogeneous structure of lithosphere depicting different tectonic blocks. Though our results are limited to shallower depth (0-90 km), it matches well with the deeper continuation of lithospheric structure obtained in an earlier study. We observe low-velocity structure all along the Eastern Himalayas down to 70 km depth, which may be attributed to deeper roots/thicker crust developed by underthrusting of Indian plate. Parallel to this low-velocity zone lies a high-velocity zone in foredeep region, represents the Indian lithosphere. The underthrusting Indian lithosphere under the Himalayas as well as below the Indo-Burma Ranges is well reflected as a high-velocity dipping structure. The buckled up part of bending Indian plate in study region, the Shillong Plateau-Mikir Hills tectonic block, is marked as a high-velocity structure at shallower depth. The Eastern Himalayan Syntaxis, tectonic block where the two arcs meet, is identified as a high-velocity structure. The Bengal Basin, tectonic block to the south of Shillong Plateau, shows low velocity due to its thicker sediments. Based on the tomographic image, a schematic model is presented to elucidate the structure and geodynamics of Indian lithosphere in study region.

  17. Induced tauopathy in a novel 3D-culture model mediates neurodegenerative processes: a real-time study on biochips.

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    Diana Seidel

    Full Text Available Tauopathies including Alzheimer's disease represent one of the major health problems of aging population worldwide. Therefore, a better understanding of tau-dependent pathologies and consequently, tau-related intervention strategies is highly demanded. In recent years, several tau-focused therapies have been proposed with the aim to stop disease progression. However, to develop efficient active pharmaceutical ingredients for the broad treatment of Alzheimer's disease patients, further improvements are necessary for understanding the detailed neurodegenerative processes as well as the mechanism and side effects of potential active pharmaceutical ingredients (API in the neuronal system. In this context, there is a lack of suitable complex in vitro cell culture models recapitulating major aspects of taupathological degenerative processes in sufficient time and reproducible manner.Herewith, we describe a novel 3D SH-SY5Y cell-based, tauopathy model that shows advanced characteristics of matured neurons in comparison to monolayer cultures without the need of artificial differentiation promoting agents. Moreover, the recombinant expression of a novel highly pathologic fourfold mutated human tau variant lead to a fast and emphasized degeneration of neuritic processes. The neurodegenerative effects could be analyzed in real time and with high sensitivity using our unique microcavity array-based impedance spectroscopy measurement system. We were able to quantify a time- and concentration-dependent relative impedance decrease when Alzheimer's disease-like tau pathology was induced in the neuronal 3D cell culture model. In combination with the collected optical information, the degenerative processes within each 3D-culture could be monitored and analyzed. More strikingly, tau-specific regenerative effects caused by tau-focused active pharmaceutical ingredients could be quantitatively monitored by impedance spectroscopy.Bringing together our novel complex 3

  18. Skin equivalent tissue-engineered construct: co-cultured fibroblasts/ keratinocytes on 3D matrices of sericin hope cocoons.

    Science.gov (United States)

    Nayak, Sunita; Dey, Sancharika; Kundu, Subhas C

    2013-01-01

    The development of effective and alternative tissue-engineered skin replacements to autografts, allografts and xenografts has became a clinical requirement due to the problems related to source of donor tissue and the perceived risk of disease transmission. In the present study 3D tissue engineered construct of sericin is developed using co-culture of keratinocytes on the upper surface of the fabricated matrices and with fibroblasts on lower surface. Sericin is obtained from "Sericin Hope" silkworm of Bombyx mori mutant and is extracted from cocoons by autoclave. Porous sericin matrices are prepared by freeze dried method using genipin as crosslinker. The matrices are characterized biochemically and biophysically. The cell proliferation and viability of co-cultured fibroblasts and keratinocytes on matrices for at least 28 days are observed by live/dead assay, Alamar blue assay, and by dual fluorescent staining. The growth of the fibroblasts and keratinocytes in co-culture is correlated with the expression level of TGF-β, b-FGF and IL-8 in the cultured supernatants by enzyme-linked immunosorbent assay. The histological analysis further demonstrates a multi-layered stratified epidermal layer of uninhibited keratinocytes in co-cultured constructs. Presence of involucrin, collagen IV and the fibroblast surface protein in immuno-histochemical stained sections of co-cultured matrices indicates the significance of paracrine signaling between keratinocytes and fibroblasts in the expression of extracellular matrix protein for dermal repair. No significant amount of pro inflammatory cytokines (TNF-α, IL-1β and nitric oxide) production are evidenced when macrophages grown on the sericin matrices. The results all together depict the potentiality of sericin 3D matrices as skin equivalent tissue engineered construct in wound repair.

  19. In vitro 3-D model based on extending time of culture for studying chronological epidermis aging.

    Science.gov (United States)

    Dos Santos, Morgan; Metral, Elodie; Boher, Aurélie; Rousselle, Patricia; Thepot, Amélie; Damour, Odile

    2015-09-01

    Skin aging is a complex phenomenon in which several mechanisms operate simultaneously. Among them, intrinsic aging is a time-dependent process, which leads to gradual skin changes affecting its structure and function such as thinning down of both epidermal and dermal compartments and a flattening and fragility of the dermo-epidermal junction. Today, several approaches have been proposed for the generation of aged skin in vitro, including skin explants from aged donors and three-dimensional skin equivalent treated by aging-inducing chemical compounds or engineered with human cells isolated from aged donors. The aim of this study was to develop and validate a new in vitro model of aging based on skin equivalent demonstrating the same phenotypic changes that were observed in chronological aging. By using prolonged culture as a proxy for cellular aging, we extended to 120 days the culture time of a skin equivalent model based on collagen-glycosaminoglycan-chitosan porous polymer and engineered with human skin cells from photo-protected sites of young donors. Morphological, immunohistological and ultrastructural analysis at different time points of the culture allowed characterizing the phenotypic changes observed in our model in comparison to samples of non photo-exposed normal human skin from different ages. We firstly confirmed that long-term cultured skin equivalents are still morphologically consistent and functionally active even after 120 days of culture. However, similar to in vivo chronological skin aging a significant decrease of the epidermis thickness as well as the number of keratinocyte expressing proliferation marker Ki67 are observed in extended culture time skin equivalent. Epidermal differentiation markers loricrin, filaggrin, involucrin and transglutaminase, also strongly decreased. Ultrastructural analysis of basement membrane showed typical features of aged skin such as duplication of lamina densa and alterations of hemidesmosomes. Moreover, the

  20. Characterization of Porcine Ventral Mesencephalic Precursor Cells following Long-Term Propagation in 3D Culture

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    Pia S. Jensen

    2012-01-01

    Full Text Available The potential use of predifferentiated neural precursor cells for treatment of a neurological disorder like Parkinson’s disease combines stem cell research with previous experimental and clinical transplantation of developing dopaminergic neurons. One current obstacle is, however, the lack of ability to generate dopaminergic neurons after long-term in vitro propagation of the cells. The domestic pig is considered a useful nonprimate large animal model in neuroscience, because of a better resemblance of the larger gyrencephalic pig brain to the human brain than the commonly used brains of smaller rodents. In the present study, porcine embryonic (28–30 days, ventral mesencephalic precursor cells were isolated and propagated as free-floating neural tissue spheres in medium containing epidermal growth factor and fibroblast growth factor 2. For passaging, the tissue spheres were cut into quarters, avoiding mechanical or enzymatic dissociation in order to minimize cellular trauma and preserve intercellular contacts. Spheres were propagated for up to 237 days with analysis of cellular content and differentiation at various time points. Our study provides the first demonstration that porcine ventral mesencephalic precursor cells can be long-term propagated as neural tissue spheres, thereby providing an experimental 3D in vitro model for studies of neural precursor cells, their niche, and differentiation capacity.

  1. Trehalose effectiveness as a cryoprotectant in 2D and 3D cell cultures of human embryonic kidney cells.

    Science.gov (United States)

    Hara, Jared; Tottori, Jordan; Anders, Megan; Dadhwal, Smritee; Asuri, Prashanth; Mobed-Miremadi, Maryam

    2017-05-01

    Post cryopreservation viability of human embryonic kidney (HEK) cells under two-dimensional (2D) and three-dimensional (3D) culture conditions was studied using trehalose as the sole cryoprotective agent. An L9 (3(4)) Taguchi design was used to optimize the cryoprotection cocktail seeding process prior to slow-freezing with the specific aim of maximizing cell viability measured 7 days post thaw, using the combinatorial cell viability and in-vitro cytotoxicity WST assay. At low (200 mM) and medium (800 mM) levels of trehalose concentration, encapsulation in alginate offered a greater protection to cryopreservation. However, at the highest trehalose concentration (1200 mM) and in the absence of the pre-incubation step, there was no statistical difference at the 95% CI (p = 0.0212) between the viability of the HEK cells under 2D and 3D culture conditions estimated to be 17.9 ± 4.6% and 14.0 ± 3.6%, respectively. A parallel comparison between cryoprotective agents conducted at the optimal levels of the L9 study, using trehalose, dimethylsulfoxide and glycerol in alginate microcapsules yielded a viability of 36.0 ± 7.4% for trehalose, in average 75% higher than the results associated with the other two cell membrane-permeating compounds. In summary, the effectiveness of trehalose has been demonstrated by the fact that 3D cell cultures can readily be equilibrated with trehalose before cryopreservation, thus mitigating the cytotoxic effects of glycerol and dimethylsulfoxide.

  2. Natural interaction in Virtual Environments for Cultural Heritage: Giotto in 3D and Etruscanning study cases

    Directory of Open Access Journals (Sweden)

    Eva Pietroni

    2012-11-01

    Full Text Available A basic limit of most of VR applications created by the scientific community and reproducing cultural sites or artefacts is that they do not fire up the attention of public, in comparison with the great potentialities of VR system for cultural transmission: they are often lacking in emotional storytelling and difficult to manage. An important factor is the need of more natural and simple interfaces, especially for applications hosted inside museums. Starting from our experience in this domain, we propose new metaphors of narration and paradigm of interaction based on natural interfaces (body movements, presenting three study cases: “The Rule confirmation: virtual experience among Giotto's characters”, “Etruscanning3D”, “Virtual Exploration of the ancient Pharmacy of S. Maria della Scaletta Hospital at Imola”.

  3. Microfluidic on-chip biomimicry for 3D cell culture: a fit-for-purpose investigation from the end user standpoint.

    Science.gov (United States)

    Liu, Ye; Gill, Elisabeth; Shery Huang, Yan Yan

    2017-06-01

    A plethora of 3D and microfluidics-based culture models have been demonstrated in the recent years with the ultimate aim to facilitate predictive in vitro models for pharmaceutical development. This article summarizes to date the progress in the microfluidics-based tissue culture models, including organ-on-a-chip and vasculature-on-a-chip. Specific focus is placed on addressing the question of what kinds of 3D culture and system complexities are deemed desirable by the biological and biomedical community. This question is addressed through analysis of a research survey to evaluate the potential use of microfluidic cell culture models among the end users. Our results showed a willingness to adopt 3D culture technology among biomedical researchers, although a significant gap still exists between the desired systems and existing 3D culture options. With these results, key challenges and future directions are highlighted.

  4. The Representation of Cultural Heritage from Traditional Drawing to 3d Survey: the Case Study of Casamary's Abbey

    Science.gov (United States)

    Canciani, M.; Saccone, M.

    2016-06-01

    In 3D survey the aspects most discussed in the scientific community are those related to the acquisition of data from integrated survey (laser scanner, photogrammetric, topographic and traditional direct), rather than those relating to the interpretation of the data. Yet in the methods of traditional representation, the data interpretation, such as that of the philological reconstruction, constitutes the most important aspect. It is therefore essential in modern systems of survey and representation, filter the information acquired. In the system, based on the integrated survey that we have adopted, the 3D object, characterized by a cloud of georeferenced points, defined but their color values, defines the core of the elaboration. It allows to carry out targeted analysis, using section planes as a tool of selection and filtering data, comparable with those of traditional drawings. In the case study of the Abbey of Casamari (Veroli), one of the most important Cistercian Settlement in Italy, the survey made for an Agreement with the Ministry of Cultural Heritage and Activities and Tourism (MiBACT) and University of RomaTre, within the project "Accessment of the sismic safety of the state museum", the reference 3D model, consisting of the superposition and geo-references data from various surveys, is the tool with which yo develop representative models comparable to traditional ones. It provides the necessary spatial environment for drawing up plans and sections with a definition such as to develop thematic analysis related to phases of construction, state of deterioration and structural features.

  5. Prognostic breast cancer signature identified from 3D culture model accurately predicts clinical outcome across independent datasets

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    Martin, Katherine J.; Patrick, Denis R.; Bissell, Mina J.; Fournier, Marcia V.

    2008-10-20

    One of the major tenets in breast cancer research is that early detection is vital for patient survival by increasing treatment options. To that end, we have previously used a novel unsupervised approach to identify a set of genes whose expression predicts prognosis of breast cancer patients. The predictive genes were selected in a well-defined three dimensional (3D) cell culture model of non-malignant human mammary epithelial cell morphogenesis as down-regulated during breast epithelial cell acinar formation and cell cycle arrest. Here we examine the ability of this gene signature (3D-signature) to predict prognosis in three independent breast cancer microarray datasets having 295, 286, and 118 samples, respectively. Our results show that the 3D-signature accurately predicts prognosis in three unrelated patient datasets. At 10 years, the probability of positive outcome was 52, 51, and 47 percent in the group with a poor-prognosis signature and 91, 75, and 71 percent in the group with a good-prognosis signature for the three datasets, respectively (Kaplan-Meier survival analysis, p<0.05). Hazard ratios for poor outcome were 5.5 (95% CI 3.0 to 12.2, p<0.0001), 2.4 (95% CI 1.6 to 3.6, p<0.0001) and 1.9 (95% CI 1.1 to 3.2, p = 0.016) and remained significant for the two larger datasets when corrected for estrogen receptor (ER) status. Hence the 3D-signature accurately predicts breast cancer outcome in both ER-positive and ER-negative tumors, though individual genes differed in their prognostic ability in the two subtypes. Genes that were prognostic in ER+ patients are AURKA, CEP55, RRM2, EPHA2, FGFBP1, and VRK1, while genes prognostic in ER patients include ACTB, FOXM1 and SERPINE2 (Kaplan-Meier p<0.05). Multivariable Cox regression analysis in the largest dataset showed that the 3D-signature was a strong independent factor in predicting breast cancer outcome. The 3D-signature accurately predicts breast cancer outcome across multiple datasets and holds prognostic

  6. Target detect system in 3D using vision apply on plant reproduction by tissue culture

    Science.gov (United States)

    Vazquez Rueda, Martin G.; Hahn, Federico

    2001-03-01

    This paper presents the preliminary results for a system in tree dimension that use a system vision to manipulate plants in a tissue culture process. The system is able to estimate the position of the plant in the work area, first calculate the position and send information to the mechanical system, and recalculate the position again, and if it is necessary, repositioning the mechanical system, using an neural system to improve the location of the plant. The system use only the system vision to sense the position and control loop using a neural system to detect the target and positioning the mechanical system, the results are compared with an open loop system.

  7. Prompt gamma-ray 3D-imaging for cultural heritage purposes

    Energy Technology Data Exchange (ETDEWEB)

    Schulze, Ralf

    2010-07-01

    The development of new, and the enhancement of existing element-sensitive imaging methods utilizing neutrons of different energy regions was the aim of the European ANCIENT CHARM project. During the present work the setup for Prompt Gamma-ray Activation Analysis (PGAA) at the research reactor FRM2 in Garching near Munich was modified to enable the spatial mapping of elemental abundances in the analysed samples. Because the PGAA setup at FRM2 was under construction at the beginning of the project first tests and the development of calibration and measurement procedures for the new imaging method were done by the PGAA group at the Budapest Research Reactor in cooperation with the Institute for Nuclear Physics of the University of Cologne. Due to the higher neutron flux at the PGAA setup at FRM2 the equipment was transferred from the Budapest Research Reactor to FRM2 after the PGAA setup at FRM2 started its regular operation. After further optimizations and the characterization of the setup, measurements were started on replicas of real archaeological objects before several measurements on real objects were performed and analysed. Several measurement configurations were applied. Additional to 2D and 3D imaging measurements a new application for the measurement of thin surface layers in the order of a few 100 {mu}m was developed. For the quantitative analysis of elemental distributions the exact knowledge of the neutron flux at each measured position in the analysed sample has to be known. Based on the well-established cold Neutron Tomography (NT) method a method and software have been developed, which enables the calculation of the neutron flux inside samples with the map of attenuation properties obtained through NT. A new data acquisition system was developed for the regular operation of the PGAA setup at FRM2, which supports traditional bulk PGAA measurements as well as measurements in the new imaging configuration. The high automation of the system allows a

  8. 3D rotating wall vessel and 2D cell culture of four veterinary virus pathogens: A comparison of virus yields, portions of infectious particles and virus growth curves.

    Science.gov (United States)

    Malenovská, Hana

    2016-02-01

    Only very few comparative studies have been performed that evaluate general trends of virus growth under 3D in comparison with 2D cell culture conditions. The aim of this study was to investigate differences when four animal viruses are cultured in 2D and 3D. Suid herpesvirus 1 (SuHV-1), Vesicular stomatitis virus (VSIV), Bovine adenovirus (BAdV) and Bovine parainfluenza 3 virus (BPIV-3) were cultivated in 3D rotating wall vessels (RWVs) and conventional 2D cultures. The production of virus particles, the portion of infectious particles, and the infectious growth curves were compared. For all viruses, the production of virus particles (related to cell density), including the non-infectious ones, was lower in 3D than in 2D culture. The production of only infectious particles was significantly lower in BAdV and BPIV-3 in 3D cultures in relation to cell density. The two cultivation approaches resulted in significantly different virus particle-to-TCID50 ratios in three of the four viruses: lower in SuHV-1 and BPIV-3 and higher in BAdV in 3D culture. The infectious virus growth rates were not significantly different in all viruses. Although 3D RWV culture resulted in lower production of virus particles compared to 2D systems, the portion of infectious particles was higher for some viruses.

  9. Preparation, characterization, and silanization of 3D microporous PDMS structure with properly sized pores for endothelial cell culture.

    Science.gov (United States)

    Zargar, Reyhaneh; Nourmohammadi, Jhamak; Amoabediny, Ghassem

    2016-01-01

    Nowadays, application of porous polydimethylsiloxane (PDMS) structure in biomedical is becoming widespread, and many methods have been established to create such structure. Although the pores created through these methods are mostly developed on the outer surface of PDMS membrane, this study offers a simple and cost-efficient technique for creating three-dimensional (3D) microporous PDMS structure with appropriate pore size for endothelial cell culture. In this study, combination of gas foaming and particulate leaching methods, with NaHCO3 as effervescent salt and NaCl as progen are used to form a 3D PDMS sponge. The in situ chemical reaction between NaHCO3 and HCl resulted in the formation of small pores and channels. Moreover, soaking the samples in HCl solution temporarily improved the hydrophilicity of PDMS, which then facilitated the penetration of water for further leaching of NaCl. The surface chemical modification process was performed by (3-aminopropyl)triethoxysilane to culture endothelial cells on porous PDMS matrix. The results are an indication of positive response of endothelial cells to the fabricated PDMS sponge. Because of simplicity and practicality of this method for preparing PDMS sponge with appropriate pore size and biological properties, the fabricated matrix can perfectly be applied to future studies in blood-contacting devices. © 2015 International Union of Biochemistry and Molecular Biology, Inc.

  10. Low cost production of 3D-printed devices and electrostimulation chambers for the culture of primary neurons.

    Science.gov (United States)

    Wardyn, Joanna D; Sanderson, Chris; Swan, Laura E; Stagi, Massimiliano

    2015-08-15

    The analysis of primary neurons is a basic requirement for many areas of neurobiology. However, the range of commercial systems available for culturing primary neurons is functionally limiting, and the expense of these devices is a barrier to both exploratory and large-scale studies. This is especially relevant as primary neurons often require unusual geometries and specialised coatings for optimum growth. Fortunately, the recent revolution in 3D printing offers the possibility to generate customised devices, which can support neuronal growth and constrain neurons in defined paths, thereby enabling many aspects of neuronal physiology to be studied with relative ease. In this article, we provide a detailed description of the system hardware and software required to produce affordable 3D-printed culture devices, which are also compatible with live-cell imaging. In addition, we also describe how to use these devices to grow and stimulate neurons within geometrically constrained compartments and provide examples to illustrate the practical utility and potential that these protocols offer for many aspects of experimental neurobiology.

  11. Interactions of Pluronic nanocarriers with 2D and 3D cell cultures: Effects of PEO block length and aggregation state.

    Science.gov (United States)

    Arranja, Alexandra; Denkova, Antonia G; Morawska, Karolina; Waton, Gilles; van Vlierberghe, Sandra; Dubruel, Peter; Schosseler, François; Mendes, Eduardo

    2016-02-28

    This work reveals how the physicochemical properties of Pluronic block copolymers influence significantly their interactions with cancer cells, whether in monolayer or spheroid cultures, and how different clinical applications can be foreseen. Two-dimensional (2D) and three-dimensional (3D) cell culture models were used to investigate the interactions of Pluronic carriers with different PEO block length and aggregation state (unimers versus cross-linked micelles) in HeLa and U87 cancer cells. Stabilized micelles of Pluronic P94 or F127 were obtained by polymerization of a crosslinking agent in the micelles hydrophobic core. Nanocarriers were functionalized with a fluorescent probe for visualization, and with a chelator for radiolabeling with Indium-111 and gamma-quantification. The 2D cell models revealed that the internalization pathways and ultimate cellular localization of the Pluronic nanocarriers depended largely on both the PEO block size and aggregation state of the copolymers. The smaller P94 unimers with an average radius of 2.1nm and the shortest PEO block mass (1100gmol(-1)) displayed the highest cellular uptake and retention. 3D tumor spheroids were used to assess the penetration capacity and toxicity potential of the nanocarriers. Results showed that cross-linked F127 micelles were more efficiently delivered across the tumor spheroids, and the penetration depth depends mostly on the transcellular transport of the carriers. The Pluronic P94-based carriers with the shortest PEO block length induced spheroid toxicity, which was significantly influenced by the spheroid cellular type.

  12. Validation of an in vitro 3D bone culture model with perfused and mechanically stressed ceramic scaffold

    Directory of Open Access Journals (Sweden)

    G Bouet

    2015-05-01

    Full Text Available An engineered three dimensional (3D in vitro cell culture system was designed with the goal of inducing and controlling in vitro osteogenesis in a reproducible manner under conditions more similar to the in vivo bone microenvironment than traditional two-dimensional (2D models. This bioreactor allows efficient mechanical loading and perfusion of an original cubic calcium phosphate bioceramic of highly controlled composition and structure. This bioceramic comprises an internal portion containing homogeneously interconnected macropores surrounded by a dense layer, which minimises fluid flow bypass around the scaffold. This dense and flat layer permits the application of a homogeneous loading on the bioceramic while also enhancing its mechanical strength. Numerical modelling of constraints shows that the system provides direct mechanical stimulation of cells within the scaffold. Experimental results establish that under perfusion at a steady flow of 2 µL/min, corresponding to 3 ≤ Medium velocity ≤ 23 µm/s, mouse calvarial cells grow and differentiate as osteoblasts in a reproducible manner, and lay down a mineralised matrix. Moreover, cells respond to mechanical loading by increasing C-fos expression, which demonstrates the effective mechanical stimulation of the culture within the scaffold. In summary, we provide a “proof-of-concept” for osteoblastic cell culture in a controlled 3D culture system under perfusion and mechanical loading. This model will be a tool to analyse bone cell functions in vivo, and will provide a bench testing system for the clinical assessment of bioactive bone-targeting molecules under load.

  13. Enhanced penetration into 3D cell culture using two and three layered gold nanoparticles

    Directory of Open Access Journals (Sweden)

    Engl

    2013-10-01

    Full Text Available Christopher G England,1 Thomas Priest,2 Guandong Zhang,2 Xinghua Sun,2 Dhruvinkumar N Patel,2 Lacey R McNally,3,4 Victor van Berkel,4,5 André M Gobin,2 Hermann B Frieboes1,2,41Department of Pharmacology and Toxicology, 2Department of Bioengineering, 3Department of Medicine, 4James Graham Brown Cancer Center, 5Department of Surgery, University of Louisville, KY, USAAbstract: Nano-scale particles sized 10–400 nm administered systemically preferentially extravasate from tumor vasculature due to the enhanced permeability and retention effect. Therapeutic success remains elusive, however, because of inhomogeneous particle distribution within tumor tissue. Insufficient tumor vascularization limits particle transport and also results in avascular hypoxic regions with non-proliferating cells, which can regenerate tissue after nanoparticle-delivered cytotoxicity or thermal ablation. Nanoparticle surface modifications provide for increasing tumor targeting and uptake while decreasing immunogenicity and toxicity. Herein, we created novel two layer gold-nanoshell particles coated with alkanethiol and phosphatidylcholine, and three layer nanoshells additionally coated with high-density-lipoprotein. We hypothesize that these particles have enhanced penetration into 3-dimensional cell cultures modeling avascular tissue when compared to standard poly(ethylene glycol (PEG-coated nanoshells. Particle uptake and distribution in liver, lung, and pancreatic tumor cell cultures were evaluated using silver-enhancement staining and hyperspectral imaging with dark field microscopy. Two layer nanoshells exhibited significantly higher uptake compared to PEGylated nanoshells. This multilayer formulation may help overcome transport barriers presented by tumor vasculature, and could be further investigated in vivo as a platform for targeted cancer therapies.Keywords: cancer nanotherapy, tumor hypoxia, nanovector transport

  14. SEE-THROUGH IMAGING OF LASER-SCANNED 3D CULTURAL HERITAGE OBJECTS BASED ON STOCHASTIC RENDERING OF LARGE-SCALE POINT CLOUDS

    OpenAIRE

    Tanaka, S.; Hasegawa, K.; Okamoto, N.; R. Umegaki; Wang, S; M. Uemura(Hiroshima Astrophysical Science Center, Hiroshima University); Okamoto, A; Koyamada, K.

    2016-01-01

    We propose a method for the precise 3D see-through imaging, or transparent visualization, of the large-scale and complex point clouds acquired via the laser scanning of 3D cultural heritage objects. Our method is based on a stochastic algorithm and directly uses the 3D points, which are acquired using a laser scanner, as the rendering primitives. This method achieves the correct depth feel without requiring depth sorting of the rendering primitives along the line of sight. Eliminatin...

  15. 3D Spheroid Culture Enhances the Expression of Antifibrotic Factors in Human Adipose-Derived MSCs and Improves Their Therapeutic Effects on Hepatic Fibrosis

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    Xuan Zhang

    2016-01-01

    Full Text Available Three-dimensional (3D cell culture has been reported to increase the therapeutic potentials of mesenchymal stem cells (MSCs. However, the action mechanisms of 3D MSCs vary greatly and are far from being thoroughly investigated. In this study, we aimed to investigate the therapeutic effects of 3D spheroids of human adipose-derived MSCs for hepatic fibrosis. Our results showed that 3D culture enhanced the expression of antifibrotic factors by MSCs, including insulin growth factor 1 (IGF-1, interleukin-6 (IL-6, and hepatocyte growth factor (HGF. In vitro studies indicated conditioned medium of 3D cultured MSCs protected hepatocytes from cell injury and apoptosis more effectively compared with 2D cultured cells. More importantly, when transplanted into model mice with hepatic fibrosis, 3D spheroids of MSCs were more beneficial in ameliorating hepatic fibrosis and improving liver function than 2D cultured cells. Therefore, the 3D culture strategy improved the therapeutic effects of MSCs and might be promising for treatment of hepatic fibrosis.

  16. 3DMADMAC|SPECTRAL: Hardware and Software Solution for Integrated Digitization of 3D Shape, Multispectral Color and BRDF for Cultural Heritage Documentation

    Directory of Open Access Journals (Sweden)

    Robert Sitnik

    2011-12-01

    Full Text Available In this article a new 3D measurement system along with the study on 3D printing technology is presented from the perspective of quality of reproduction. In the first part of the paper the 3DMADMAC|SPECTRAL system which integrates 3D shape with additional color and angular reflectance measurement capabilities is presented (see Figure 1. The shape measurement system is based on structured light projection with the use of a DLP projector. The 3D shape measurement method is based on sinusoidal fringes and Gray codes projection. Color is being measured using multispectral images with a set of interference filters to separate spectral channels. Additionally the set up includes an array of compact light sources for measuring angular reflectance based on image analysis and 3D data processing. All three components of the integrated system use the same greyscale camera as a detector. The purpose of the system is to obtain complete information about shape, color and reflectance characteristic of mea sured surface, especially for cultural heritage objects - in order to create high quality 3D documentation. In the second part of the paper the 3D printing technology will be tested on real measured cultural heritage objects. Tests allow to assess measurement and color accuracy of reproduction by selected 3D printing technology and shed some light on how current 3D printing technology can be applied into cultural heritage.

  17. Bile canaliculi formation and biliary transport in 3D sandwich-cultured hepatocytes in dependence of the extracellular matrix composition.

    Science.gov (United States)

    Deharde, Daniela; Schneider, Christin; Hiller, Thomas; Fischer, Nicolas; Kegel, Victoria; Lübberstedt, Marc; Freyer, Nora; Hengstler, Jan G; Andersson, Tommy B; Seehofer, Daniel; Pratschke, Johann; Zeilinger, Katrin; Damm, Georg

    2016-10-01

    Primary human hepatocytes (PHH) are still considered as gold standard for investigation of in vitro metabolism and hepatotoxicity in pharmaceutical research. It has been shown that the three-dimensional (3D) cultivation of PHH in a sandwich configuration between two layers of extracellular matrix (ECM) enables the hepatocytes to adhere three dimensionally leading to formation of in vivo like cell-cell contacts and cell-matrix interactions. The aim of the present study was to investigate the influence of different ECM compositions on morphology, cellular arrangement and bile canaliculi formation as well as bile excretion processes in PHH sandwich cultures systematically. Freshly isolated PHH were cultured for 6 days between two ECM layers made of collagen and/or Matrigel in four different combinations. The cultures were investigated by phase contrast microscopy and immunofluorescence analysis with respect to cell-cell connections, repolarization as well as bile canaliculi formation. The influence of the ECM composition on cell activity and viability was measured using the XTT assay and a fluorescent dead or alive assay. Finally, the bile canalicular transport was analyzed by live cell imaging to monitor the secretion and accumulation of the fluorescent substance CDF in bile canaliculi. Using collagen and Matrigel in different compositions in sandwich cultures of hepatocytes, we observed differences in morphology, cellular arrangement and cell activity of PHH in dependence of the ECM composition. Sandwich-cultured hepatocytes with an underlay of collagen seem to represent the best in vivo tissue architecture in terms of formation of trabecular cell arrangement. Cultures overlaid with collagen were characterized by the formation of abundant bile canaliculi, while the bile canaliculi network in hepatocytes cultured on a layer of Matrigel and overlaid with collagen showed the most branched and stable canalicular network. All cultures showed a time-dependent leakage of

  18. 3D Culture of Chondrocytes in Gelatin Hydrogels with Different Stiffness

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    Xiaomeng Li

    2016-07-01

    Full Text Available Gelatin hydrogels can mimic the microenvironments of natural tissues and encapsulate cells homogeneously, which makes them attractive for cartilage tissue engineering. Both the mechanical and biochemical properties of hydrogels can affect the phenotype of chondrocytes. However, the influence of each property on chondrocyte phenotype is unclear due to the difficulty in separating the roles of these properties. In this study, we aimed to study the influence of hydrogel stiffness on chondrocyte phenotype while excluding the role of biochemical factors, such as adhesion site density in the hydrogels. By altering the degree of methacryloyl functionalization, gelatin hydrogels with different stiffnesses of 3.8, 17.1, and 29.9 kPa Young’s modulus were prepared from the same concentration of gelatin methacryloyl (GelMA macromers. Bovine articular chondrocytes were encapsulated in the hydrogels and cultured for 14 days. The influence of hydrogel stiffness on the cell behaviors including cell viability, cell morphology, and maintenance of chondrogenic phenotype was evaluated. GelMA hydrogels with high stiffness (29.9 kPa showed the best results on maintaining chondrogenic phenotype. These results will be useful for the design and preparation of scaffolds for cartilage tissue engineering.

  19. Time lapse investigation of antibiotic susceptibility using a microfluidic linear gradient 3D culture device.

    Science.gov (United States)

    Hou, Zining; An, Yu; Hjort, Karin; Hjort, Klas; Sandegren, Linus; Wu, Zhigang

    2014-09-01

    This study reports a novel approach to quantitatively investigate the antibacterial effect of antibiotics on bacteria using a three-dimensional microfluidic culture device. In particular, our approach is suitable for studying the pharmacodynamics effects of antibiotics on bacterial cells temporally and with a continuous range of concentrations in a single experiment. The responses of bacterial cells to a linear concentration gradient of antibiotics were observed using time-lapse photography, by encapsulating bacterial cells in an agarose-based gel located in a commercially available microfluidics chamber. This approach generates dynamic information with high resolution, in a single operation, e.g., growth curves and antibiotic pharmacodynamics, in a well-controlled environment. No pre-labelling of the cells is needed and therefore any bacterial sample can be tested in this setup. It also provides static information comparable to that of standard techniques for measuring minimum inhibitory concentration (MIC). Five antibiotics with different mechanisms were analysed against wild-type Escherichia coli, Staphylococcus aureus and Salmonella Typhimurium. The entire process, including data analysis, took 2.5-4 h and from the same analysis, high-resolution growth curves were obtained. As a proof of principle, a pharmacodynamic model of streptomycin against Salmonella Typhimurium was built based on the maximal effect model, which agreed well with the experimental results. Our approach has the potential to be a simple and flexible solution to study responding behaviours of microbial cells under different selection pressures both temporally and in a range of concentrations.

  20. Multiplex profiling of cellular invasion in 3D cell culture models.

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    Gerald Burgstaller

    Full Text Available To-date, most invasion or migration assays use a modified Boyden chamber-like design to assess migration as single-cell or scratch assays on coated or uncoated planar plastic surfaces. Here, we describe a 96-well microplate-based, high-content, three-dimensional cell culture assay capable of assessing invasion dynamics and molecular signatures thereof. On applying our invasion assay, we were able to demonstrate significant effects on the invasion capacity of fibroblast cell lines, as well as primary lung fibroblasts. Administration of epidermal growth factor resulted in a substantial increase of cellular invasion, thus making this technique suitable for high-throughput pharmacological screening of novel compounds regulating invasive and migratory pathways of primary cells. Our assay also correlates cellular invasiveness to molecular events. Thus, we argue of having developed a powerful and versatile toolbox for an extensive profiling of invasive cells in a 96-well format. This will have a major impact on research in disease areas like fibrosis, metastatic cancers, or chronic inflammatory states.

  1. Cell therapy, 3D culture systems and tissue engineering for cardiac regeneration.

    Science.gov (United States)

    Emmert, Maximilian Y; Hitchcock, Robert W; Hoerstrup, Simon P

    2014-04-01

    Ischemic Heart Disease (IHD) still represents the "Number One Killer" worldwide accounting for the death of numerous patients. However the capacity for self-regeneration of the adult heart is very limited and the loss of cardiomyocytes in the infarcted heart leads to continuous adverse cardiac-remodeling which often leads to heart-failure (HF). The concept of regenerative medicine comprising cell-based therapies, bio-engineering technologies and hybrid solutions has been proposed as a promising next-generation approach to address IHD and HF. Numerous strategies are under investigation evaluating the potential of regenerative medicine on the failing myocardium including classical cell-therapy concepts, three-dimensional culture techniques and tissue-engineering approaches. While most of these regenerative strategies have shown great potential in experimental studies, the translation into a clinical setting has either been limited or too rapid leaving many key questions unanswered. This review summarizes the current state-of-the-art, important challenges and future research directions as to regenerative approaches addressing IHD and resulting HF.

  2. How linguistic and cultural forces shape conceptions of time: English and Mandarin time in 3D.

    Science.gov (United States)

    Fuhrman, Orly; McCormick, Kelly; Chen, Eva; Jiang, Heidi; Shu, Dingfang; Mao, Shuaimei; Boroditsky, Lera

    2011-01-01

    In this paper we examine how English and Mandarin speakers think about time, and we test how the patterns of thinking in the two groups relate to patterns in linguistic and cultural experience. In Mandarin, vertical spatial metaphors are used more frequently to talk about time than they are in English; English relies primarily on horizontal terms. We present results from two tasks comparing English and Mandarin speakers' temporal reasoning. The tasks measure how people spatialize time in three-dimensional space, including the sagittal (front/back), transverse (left/right), and vertical (up/down) axes. Results of Experiment 1 show that people automatically create spatial representations in the course of temporal reasoning, and these implicit spatializations differ in accordance with patterns in language, even in a non-linguistic task. Both groups showed evidence of a left-to-right representation of time, in accordance with writing direction, but only Mandarin speakers showed a vertical top-to-bottom pattern for time (congruent with vertical spatiotemporal metaphors in Mandarin). Results of Experiment 2 confirm and extend these findings, showing that bilinguals' representations of time depend on both long-term and proximal aspects of language experience. Participants who were more proficient in Mandarin were more likely to arrange time vertically (an effect of previous language experience). Further, bilinguals were more likely to arrange time vertically when they were tested in Mandarin than when they were tested in English (an effect of immediate linguistic context). Copyright © 2011 Cognitive Science Society, Inc.

  3. Photopatterning of hydrogel scaffolds coupled to filter materials using stereolithography for perfused 3D culture of hepatocytes.

    Science.gov (United States)

    Neiman, Jaclyn A Shepard; Raman, Ritu; Chan, Vincent; Rhoads, Mary G; Raredon, Micha Sam B; Velazquez, Jeremy J; Dyer, Rachel L; Bashir, Rashid; Hammond, Paula T; Griffith, Linda G

    2015-04-01

    In vitro models that recapitulate the liver's structural and functional complexity could prolong hepatocellular viability and function to improve platforms for drug toxicity studies and understanding liver pathophysiology. Here, stereolithography (SLA) was employed to fabricate hydrogel scaffolds with open channels designed for post-seeding and perfused culture of primary hepatocytes that form 3D structures in a bioreactor. Photopolymerizable polyethylene glycol-based hydrogels were fabricated coupled to chemically activated, commercially available filters (polycarbonate and polyvinylidene fluoride) using a chemistry that permitted cell viability, and was robust enough to withstand perfused culture of up to 1 µL/s for at least 7 days. SLA energy dose, photoinitiator concentrations, and pretreatment conditions were screened to determine conditions that maximized cell viability and hydrogel bonding to the filter. Multiple open channel geometries were readily achieved, and included ellipses and rectangles. Rectangular open channels employed for subsequent studies had final dimensions on the order of 350 µm by 850 µm. Cell seeding densities and flow rates that promoted cell viability were determined. Perfused culture of primary hepatocytes in hydrogel scaffolds in the presence of soluble epidermal growth factor (EGF) prolonged the maintenance of albumin production throughout the 7-day culture relative to 2D controls. This technique of bonding hydrogel scaffolds can be employed to fabricate soft scaffolds for a number of bioreactor configurations and applications.

  4. Effects of intermittent hydrostatic pressure magnitude on the chondrogenesis of MSCs without biochemical agents under 3D co-culture.

    Science.gov (United States)

    Jeong, Jae Young; Park, So Hee; Shin, Ji Won; Kang, Yun Gyeong; Han, Ki-Ho; Shin, Jung-Woog

    2012-11-01

    Without using biochemical agents, in this study, we sought to investigate the potential of controlling the differentiation of mesenchymal stem cells (MSCs) into a specific cell type through the use of 3D co-culturing and mechanical stimuli. MSCs and primary cultured chondrocytes were separately encapsulated into alginate beads, and the two types of beads were separated by a membrane. For the investigation a computer-controllable bioreactor was designed and used to engage intermittent hydrostatic pressure (IHP). Five different magnitudes (0.20, 0.10, 0.05, 0.02 MPa and no stimulation) of IHP were applied. The stimulation pattern was the same for all groups: 2 h/day for 7 days starting at 24 h after seeding; 2 and 15 min cycles of stimulating and resting, respectively. Biochemical (DNA and GAG contents), histological (Alcian blue), and RT-PCR (Col II, SOX9, AGC) analyses were performed on days 1, 5, 10, and 20. The results from these analyses showed that stimulation with higher magnitudes of IHP (≥0.10 MPa) were more effective on the proliferation and differentiation of co-cultured MSCs. Together, these data demonstrate the potential of using mechanical stimulation and co-culturing for the proliferation and differentiation of MSCs, even without biochemical agents.

  5. Genotoxicity assessment of reactive and disperse textile dyes using human dermal equivalent (3D cell culture system).

    Science.gov (United States)

    Leme, Daniela Morais; Primo, Fernando Lucas; Gobo, Graciely Gomides; da Costa, Cleber Rafael Vieira; Tedesco, Antonio Claudio; de Oliveira, Danielle Palma

    2015-01-01

    Thousands of dyes are marketed daily for different purposes, including textile dyeing. However, there are several studies reporting attributing to dyes deleterious human effects such as DNA damage. Humans may be exposed to toxic dyes through either ingestion of contaminated waters or dermal contact with colored garments. With respect to dermal exposure, human skin equivalents are promising tools to assess in vitro genotoxicity of dermally applied chemicals using a three-dimensional (3D) model to mimic tissue behavior. This study investigated the sensitivity of an in-house human dermal equivalent (DE) for detecting genotoxicity of textile dyes. Two azo (reactive green 19 [RG19] and disperse red 1[DR1]) dyes and one anthraquinone (reactive blue 2 [RB2]) dye were analyzed. RG19 was genotoxic for DE in a dose-responsive manner, whereas RB2 and DR1 were nongenotoxic under the conditions tested. These findings are not in agreement with previous genotoxicological assessment of these dyes carried out using two-dimensional (2D) cell cultures, which showed that DR1 was genotoxic in human hepatoma cells (HepG2) and RG19 was nongenotoxic for normal human dermal fibroblasts (NHDF). These discrepant results probably may be due to differences between metabolic activities of each cell type (organ-specific genotoxicity, HepG2 and fibroblasts) and the test setup systems used in each study (fibroblasts cultured at 2D and three-dimensional [3D] culture systems). Genotoxicological assessment of textile dyes in context of organ-specific genotoxicity and using in vitro models that more closely resemble in vivo tissue architecture and physiology may provide more reliable estimates of genotoxic potential of these chemicals.

  6. A fully defined and scalable 3D culture system for human pluripotent stem cell expansion and differentiation

    Science.gov (United States)

    Lei, Yuguo; Schaffer, David V.

    2013-12-01

    Human pluripotent stem cells (hPSCs), including human embryonic stem cells and induced pluripotent stem cells, are promising for numerous biomedical applications, such as cell replacement therapies, tissue and whole-organ engineering, and high-throughput pharmacology and toxicology screening. Each of these applications requires large numbers of cells of high quality; however, the scalable expansion and differentiation of hPSCs, especially for clinical utilization, remains a challenge. We report a simple, defined, efficient, scalable, and good manufacturing practice-compatible 3D culture system for hPSC expansion and differentiation. It employs a thermoresponsive hydrogel that combines easy manipulation and completely defined conditions, free of any human- or animal-derived factors, and entailing only recombinant protein factors. Under an optimized protocol, the 3D system enables long-term, serial expansion of multiple hPSCs lines with a high expansion rate (∼20-fold per 5-d passage, for a 1072-fold expansion over 280 d), yield (∼2.0 × 107 cells per mL of hydrogel), and purity (∼95% Oct4+), even with single-cell inoculation, all of which offer considerable advantages relative to current approaches. Moreover, the system enabled 3D directed differentiation of hPSCs into multiple lineages, including dopaminergic neuron progenitors with a yield of ∼8 × 107 dopaminergic progenitors per mL of hydrogel and ∼80-fold expansion by the end of a 15-d derivation. This versatile system may be useful at numerous scales, from basic biological investigation to clinical development.

  7. The influence of nutrient supply and cell density on the growth and survival of intervertebral disc cells in 3D culture

    Directory of Open Access Journals (Sweden)

    S Stephan

    2011-09-01

    Full Text Available The adult human intervertebral disc (IVD is normally avascular. Changes to the extracellular matrix in degenerative disc disease may promote vascularisation and subsequently alter cell nutrition and disc homeostasis. This study examines the influence of cell density and the presence of glucose and serum on the proliferation and survival of IVD cells in 3D culture.Bovine nucleus pulposus (NP cells were seeded at a range of cell densities (1.25 x105-106 cells/mL and cultured in alginate beads under standard culture conditions (with 3.15 g/L glucose and 10 % serum, or without glucose and/or 20 % serum. Cell proliferation, apoptosis and cell senescence were examined after 8 days in culture.Under standard culture conditions, NP cell proliferation and cluster formation was inversely related to cell seeding density, whilst the number of apoptotic cells and enucleated “ghost” cells was positively correlated to cell seeding density. Increasing serum levels from 10 % to 20 % was associated with increased cluster size and also an increased prevalence of apoptotic cells within clusters. Omitting glucose produced even larger clusters and also more apoptotic and senescent cells. These studies demonstrate that NP cell growth and survival are influenced both by cell density and the availability of serum or nutrients, such as glucose. The observation of clustered, senescent, apoptotic or “ghost” cells in vitro suggests that environmental factors may influence the formation of these phenotypes that have been previously reported in vivo. Hence this study has implications for both our understanding of degenerative disc disease and also cell-based therapy using cells cultured in vitro.

  8. See-Through Imaging of Laser-Scanned 3d Cultural Heritage Objects Based on Stochastic Rendering of Large-Scale Point Clouds

    Science.gov (United States)

    Tanaka, S.; Hasegawa, K.; Okamoto, N.; Umegaki, R.; Wang, S.; Uemura, M.; Okamoto, A.; Koyamada, K.

    2016-06-01

    We propose a method for the precise 3D see-through imaging, or transparent visualization, of the large-scale and complex point clouds acquired via the laser scanning of 3D cultural heritage objects. Our method is based on a stochastic algorithm and directly uses the 3D points, which are acquired using a laser scanner, as the rendering primitives. This method achieves the correct depth feel without requiring depth sorting of the rendering primitives along the line of sight. Eliminating this need allows us to avoid long computation times when creating natural and precise 3D see-through views of laser-scanned cultural heritage objects. The opacity of each laser-scanned object is also flexibly controllable. For a laser-scanned point cloud consisting of more than 107 or 108 3D points, the pre-processing requires only a few minutes, and the rendering can be executed at interactive frame rates. Our method enables the creation of cumulative 3D see-through images of time-series laser-scanned data. It also offers the possibility of fused visualization for observing a laser-scanned object behind a transparent high-quality photographic image placed in the 3D scene. We demonstrate the effectiveness of our method by applying it to festival floats of high cultural value. These festival floats have complex outer and inner 3D structures and are suitable for see-through imaging.

  9. Development of a 3D Tissue Culture-Based High-Content Screening Platform That Uses Phenotypic Profiling to Discriminate Selective Inhibitors of Receptor Tyrosine Kinases.

    Science.gov (United States)

    Booij, Tijmen H; Klop, Maarten J D; Yan, Kuan; Szántai-Kis, Csaba; Szokol, Balint; Orfi, Laszlo; van de Water, Bob; Keri, Gyorgy; Price, Leo S

    2016-10-01

    3D tissue cultures provide a more physiologically relevant context for the screening of compounds, compared with 2D cell cultures. Cells cultured in 3D hydrogels also show complex phenotypes, increasing the scope for phenotypic profiling. Here we describe a high-content screening platform that uses invasive human prostate cancer cells cultured in 3D in standard 384-well assay plates to study the activity of potential therapeutic small molecules and antibody biologics. Image analysis tools were developed to process 3D image data to measure over 800 phenotypic parameters. Multiparametric analysis was used to evaluate the effect of compounds on tissue morphology. We applied this screening platform to measure the activity and selectivity of inhibitors of the c-Met and epidermal growth factor (EGF) receptor (EGFR) tyrosine kinases in 3D cultured prostate carcinoma cells. c-Met and EGFR activity was quantified based on the phenotypic profiles induced by their respective ligands, hepatocyte growth factor and EGF. The screening method was applied to a novel collection of 80 putative inhibitors of c-Met and EGFR. Compounds were identified that induced phenotypic profiles indicative of selective inhibition of c-Met, EGFR, or bispecific inhibition of both targets. In conclusion, we describe a fully scalable high-content screening platform that uses phenotypic profiling to discriminate selective and nonselective (off-target) inhibitors in a physiologically relevant 3D cell culture setting.

  10. THE REPRESENTATION OF CULTURAL HERITAGE FROM TRADITIONAL DRAWING TO 3D SURVEY: THE CASE STUDY OF CASAMARY’S ABBEY

    Directory of Open Access Journals (Sweden)

    M. Canciani

    2016-06-01

    In the case study of the Abbey of Casamari (Veroli, one of the most important Cistercian Settlement in Italy, the survey made for an Agreement with the Ministry of Cultural Heritage and Activities and Tourism (MiBACT and University of RomaTre, within the project "Accessment of the sismic safety of the state museum", the reference 3D model, consisting of the superposition and geo-references data from various surveys, is the tool with which yo develop representative models comparable to traditional ones. It provides the necessary spatial environment for drawing up plans and sections with a definition such as to develop thematic analysis related to phases of construction, state of deterioration and structural features.

  11. Composite Scaffolds Containing Silk Fibroin, Gelatin, and Hydroxyapatite for Bone Tissue Regeneration and 3D Cell Culturing.

    Science.gov (United States)

    Moisenovich, M M; Arkhipova, A Yu; Orlova, A A; Drutskaya, M S; Volkova, S V; Zacharov, S E; Agapov, I I; Kirpichnikov, M P

    2014-01-01

    Three-dimensional (3D) silk fibroin scaffolds were modified with one of the major bone tissue derivatives (nano-hydroxyapatite) and/or a collagen derivative (gelatin). Adhesion and proliferation of mouse embryonic fibroblasts (MEF) within the scaffold were increased after modification with either nano-hydroxyapatite or gelatin. However, a significant increase in MEF adhesion and proliferation was observed when both additives were introduced into the scaffold. Such modified composite scaffolds provide a new and better platform to study wound healing, bone and other tissue regeneration, as well as artificial organ bioengineering. This system can further be applied to establish experimental models to study cell-substrate interactions, cell migration and other complex processes, which may be difficult to address using the conventional two-dimensional culture systems.

  12. Optimization and comparison of two different 3D culture methods to prepare cell aggregates as a bioink for organ printing.

    Science.gov (United States)

    Imani, Rana; Hojjati Emami, Shahriar; Fakhrzadeh, Hossein; Baheiraei, Nafiseh; Sharifi, Ali M

    2012-04-01

    The ultimate goal of tissue engineering is to design and fabricate functional human tissues that are similar to natural cells and are capable of regeneration. Preparation of cell aggregates is one of the important steps in 3D tissue engineering technology, particularly in organ printing. Two simple methods, hanging drop (HD) and conical tube (CT) were utilized to prepare cell aggregates. The size and viability of the aggregates obtained at different initial cell densities and pre-culture duration were compared. The proliferative ability of the cell aggregates and their ability to spread in culture plates were also investigated. In both methods, the optimum average size of the aggregates was less than 500 microm. CT aggregates were smaller than HD aggregates. 5,000 cells per drop HD aggregates showed a marked ability to attach and spread on the culture surface. The proliferative ability reduced when the initial cell density was increased. Comparing these methods, we found that the HD method having better size controlling ability as well as enhanced ability to maintain higher rates of viability, spreading, and proliferation. In conclusion, smaller HD aggregates might be a suitable choice as building blocks for making bioink particles in bioprinting technique.

  13. The microenvironment determines the breast cancer cells' phenotype: organization of MCF7 cells in 3D cultures

    Directory of Open Access Journals (Sweden)

    Soto Ana M

    2010-06-01

    Full Text Available Abstract Background Stromal-epithelial interactions mediate breast development, and the initiation and progression of breast cancer. In the present study, we developed 3-dimensional (3D in vitro models to study breast cancer tissue organization and the role of the microenvironment in phenotypic determination. Methods The human breast cancer MCF7 cells were grown alone or co-cultured with primary human breast fibroblasts. Cells were embedded in matrices containing either type I collagen or a combination of reconstituted basement membrane proteins and type I collagen. The cultures were carried out for up to 6 weeks. For every time point (1-6 weeks, the gels were fixed and processed for histology, and whole-mounted for confocal microscopy evaluation. The epithelial structures were characterized utilizing immunohistochemical techniques; their area and proliferation index were measured using computerized morphometric analysis. Statistical differences between groups were analyzed by ANOVA, Dunnett's T3 post-hoc test and chi-square. Results Most of the MCF7 cells grown alone within a collagen matrix died during the first two weeks; those that survived organized into large, round and solid clusters. The presence of fibroblasts in collagen gels reduced MCF7 cell death, induced cell polarity, and the formation of round and elongated epithelial structures containing a lumen. The addition of reconstituted basement membrane to collagen gels by itself had also survival and organizational effects on the MCF7 cells. Regardless of the presence of fibroblasts, the MCF7 cells both polarized and formed a lumen. The addition of fibroblasts to the gel containing reconstituted basement membrane and collagen induced the formation of elongated structures. Conclusions Our results indicate that a matrix containing both type I collagen and reconstituted basement membrane, and the presence of normal breast fibroblasts constitute the minimal permissive microenvironment to

  14. Investigation of Adaptive Responses in Bystander Cells in 3D Cultures Containing Tritium-Labeled and Unlabeled Normal Human Fibroblasts

    Science.gov (United States)

    Pinto, Massimo; Azzam, Edouard I.; Howell, Roger W.

    2010-01-01

    The study of radiation-induced bystander effects in normal human cells maintained in three-dimensional (3D) architecture provides more in vivo-like conditions and is relevant to human risk assessment. Linear energy transfer, dose and dose rate have been considered as critical factors in propagating radiation-induced effects. This investigation uses an in vitro 3D tissue culture model in which normal AG1522 human fibroblasts are grown in a carbon scaffold to investigate induction of a G1 arrest in bystander cells that neighbor radiolabeled cells. Cell cultures were co-pulse-labeled with [3H]deoxycytidine (3HdC) to selectively irradiate a minor fraction of cells with 1–5 keV/μm β particles and bromodeoxyuridine (BrdU) to identify the radiolabeled cells using immunofluorescence. The induction of a G1 arrest was measured specifically in unlabeled cells (i.e. bystander cells) using a flow cytometry-based version of the cumulative labeling index assay. To investigate the relationship between bystander effects and adaptive responses, cells were challenged with an acute 4 Gy γ-radiation dose after they had been kept under the bystander conditions described above for several hours, and the regulation of the radiation-induced G1 arrest was measured selectively in bystander cells. When the average dose rate in 3HdC-labeled cells (bystander effects or adaptive bystander effects were observed as measured by magnitude of the G1 arrest, micronucleus formation, or changes in mitochondrial membrane potential. Higher dose rates and/or higher LET may be required to observe stressful bystander effects in this experimental system, whereas lower dose rates and challenge doses may be required to detect adaptive bystander responses. PMID:20681788

  15. Breast epithelial tissue morphology is affected in 3D cultures by species-specific collagen-based extracellular matrix.

    Science.gov (United States)

    Dhimolea, Eugen; Soto, Ana M; Sonnenschein, Carlos

    2012-11-01

    Collagen-based gels have been widely used to determine the factors that regulate branching morphogenesis in the mammary gland. The patterns of biomechanical gradients and collagen reorganization influence the shape and orientation of epithelial structures in three-dimensional (3D) conditions. We explored in greater detail whether collagen type I fibers with distinct biomechanical and fiber-assembling properties, isolated from either bovine or rat tail tendon, differentially affected the epithelial phenotype in a tissue culture model of the human breast. Rat tail collagen fibers were densely packed into significantly longer and thicker bundles compared to those of the bovine type (average fascicle length 7.35 and 2.29 μm, respectively; p = 0.0001), indicating increased fiber alignment and biomechanical enablement in the former. MCF10A epithelial cells formed elaborated branched tubular structures in bovine but only nonbranched ducts and acini in rat tail collagen matrices. Ductal branching in bovine collagen was associated with interactions between neighboring structures mediated through packed collagen fibers; these fiber-mediated interactions were absent in rat tail collagen gels. Normal breast fibroblasts increased the final size and number of ducts only in rat tail collagen gels while not affecting branching. Our results suggest that the species of origin of collagen used in organotypic cultures may influence epithelial differentiation into alveolar or ductal structures and the patterns of epithelial branching. These observations underscore the importance of considering the species of origin and fiber alignment properties of collagen when engineering branching organs in 3D matrices and interpreting their role in the tissue phenotype.

  16. The methodology of documenting cultural heritage sites using photogrammetry, UAV, and 3D printing techniques: the case study of Asinou Church in Cyprus

    Science.gov (United States)

    Themistocleous, K.; Ioannides, M.; Agapiou, A.; Hadjimitsis, D. G.

    2015-06-01

    As the affordability, reliability and ease-of-use of Unmanned Aerial Vehicles (UAV) advances, the use of aerial surveying for cultural heritage purposes becomes a popular choice, yielding an unprecedented volume of high-resolution, geo-tagged image-sets of historical sites from above. As well, recent developments in photogrammetry technology provide a simple and cost-effective method of generating relatively accurate 3D models from 2D images. These techniques provide a set of new tools for archaeologists and cultural heritage experts to capture, store, process, share, visualise and annotate 3D models in the field. This paper focuses on the methodology used to document the cultural heritage site of Asinou Church in Cyprus using various state of the art techniques, such as UAV, photogrammetry and 3D printing. Hundreds of images of the Asinou Church were taken by a UAV with an attached high resolution, low cost camera. These photographic images were then used to create a digital 3D model and a 3D printer was used to create a physical model of the church. Such a methodology provides archaeologists and cultural heritage experts a simple and cost-effective method of generating relatively accurate 3D models from 2D images of cultural heritage sites.

  17. A 3D cell culture system: separation distance between INS-1 cell and endothelial cell monolayers co-cultured in fibrin influences INS-1 cells insulin secretion.

    Science.gov (United States)

    Sabra, Georges; Vermette, Patrick

    2013-02-01

    The aim of this study was to develop an in vitro cell culture system allowing studying the effect of separation distance between monolayers of rat insulinoma cells (INS-1) and human umbilical vein endothelial cells (HUVEC) co-cultured in fibrin over INS-1 cell insulin secretion. For this purpose, a three-dimensional (3D) cell culture chamber was designed, built using micro-fabrication techniques and validated. The co-culture was successfully carried out and the effect on INS-1 cell insulin secretion was investigated. After 48 and 72 h, INS-1 cells co-cultured with HUVEC separated by a distance of 100 µm revealed enhanced insulin secretion compared to INS-1 cells cultured alone or co-cultured with HUVEC monolayers separated by a distance of 200 µm. These results illustrate the importance of the separation distance between two cell niches for cell culture design and the possibility to further enhance the endocrine function of beta cells when this factor is considered.

  18. Influence of Matrices on 3D-Cultured Prostate Cancer Cells' Drug Response and Expression of Drug-Action Associated Proteins.

    Science.gov (United States)

    Edmondson, Rasheena; Adcock, Audrey F; Yang, Liju

    2016-01-01

    This study investigated the effects of matrix on the behaviors of 3D-cultured cells of two prostate cancer cell lines, LNCaP and DU145. Two biologically-derived matrices, Matrigel and Cultrex BME, and one synthetic matrix, the Alvetex scaffold, were used to culture the cells. The cell proliferation rate, cellular response to anti-cancer drugs, and expression levels of proteins associated with drug sensitivity/resistance were examined and compared amongst the 3D-cultured cells on the three matrices and 2D-cultured cells. The cellular responses upon treatment with two common anti-cancer drugs, Docetaxel and Rapamycin, were examined. The expressions of epidermal growth factor receptor (EGFR) and β-III tubulin in DU145 cells and p53 in LNCaP cells were examined. The results showed that the proliferation rates of cells cultured on the three matrices varied, especially between the synthetic matrix and the biologically-derived matrices. The drug responses and the expressions of drug sensitivity-associated proteins differed between cells on various matrices as well. Among the 3D cultures on the three matrices, increased expression of β-III tubulin in DU145 cells was correlated with increased resistance to Docetaxel, and decreased expression of EGFR in DU145 cells was correlated with increased sensitivity to Rapamycin. Increased expression of a p53 dimer in 3D-cultured LNCaP cells was correlated with increased resistance to Docetaxel. Collectively, the results showed that the matrix of 3D cell culture models strongly influences cellular behaviors, which highlights the imperative need to achieve standardization of 3D cell culture technology in order to be used in drug screening and cell biology studies.

  19. Probing tumor-stroma interactions and response to photodynamic therapy in a 3D pancreatic cancer-fibroblast co-culture model

    Science.gov (United States)

    Glidden, Michael D.; Massodi, Iqbal; Rizvi, Imran; Celli, Jonathan P.; Hasan, Tayyaba

    2012-02-01

    Pancreatic ductal adenocarcinoma is a lethal disease that is often unresectable by the time of diagnosis and is typically non-responsive to chemo- and radiotherapy, resulting in a five year survival of only 3%. Tumors of the pancreas are characterized by a dense fibrous stroma rich in extracellular matrix proteins, which is implicated in poor therapeutic response, though its precise roles remain poorly understood. Indeed, while the use of therapeutics that target the stroma is an emerging paradigm in the clinical management of this disease, the primary focus of such efforts is to enhance drug penetration through dense fibrous stroma and it is unclear to what extent the characteristically rigid stroma of pancreatic tumors imparts drug resistance by acting as a complex signaling partner, or merely as a physical barrier for drug delivery. Here we use 3D in vitro co-cultures of pancreatic cancer cells and normal human fibroblasts as a model system to study heterotypic interactions between these populations. Leveraging this in vitro model along with image-based methods for quantification of growth and therapeutic endpoints, we characterize these co-cultures and examine the role of verteporfin-based photodynamic therapy (PDT) for targeting tumor-fibroblast interactions in pancreatic tumors.

  20. Estimation of bisphenol A-Human toxicity by 3D cell culture arrays, high throughput alternatives to animal tests.

    Science.gov (United States)

    Lee, Dong Woo; Oh, Woo-Yeon; Yi, Sang Hyun; Ku, Bosung; Lee, Moo-Yeal; Cho, Yoon Hee; Yang, Mihi

    2016-09-30

    Bisphenol A (BPA) has been widely used for manufacturing polycarbonate plastics and epoxy resins and has been extensively tested in animals to predict human toxicity. In order to reduce the use of animals for toxicity assessment and provide further accurate information on BPA toxicity in humans, we encapsulated Hep3B human hepatoma cells in alginate and cultured them in three dimensions (3D) on a micropillar chip coupled to a panel of metabolic enzymes on a microwell chip. As a result, we were able to assess the toxicity of BPA under various metabolic enzyme conditions using a high-throughput and micro assay; sample volumes were nearly 2,000 times less than that required for a 96-well plate. We applied a total of 28 different enzymes to each chip, including 10 cytochrome P450s (CYP450s), 10 UDP-glycosyltransferases (UGTs), 3 sulfotransferases (SULTs), alcohol dehydrogenase (ADH), and aldehyde dehydrogenase 2 (ALDH2). Phase I enzyme mixtures, phase II enzyme mixtures, and a combination of phase I and phase II enzymes were also applied to the chip. BPA toxicity was higher in samples containing CYP2E1 than controls, which contained no enzymes (IC50, 184±16μM and 270±25.8μM, respectively, panimal testing for estimating BPA toxicity via human metabolic systems. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  1. Chrysotile effects on human lung cell carcinoma in culture: 3-D reconstruction and DNA quantification by image analysis

    Directory of Open Access Journals (Sweden)

    Machado-Santelli Glaucia M

    2008-06-01

    Full Text Available Abstract Background Chrysotile is considered less harmful to human health than other types of asbestos fibers. Its clearance from the lung is faster and, in comparison to amphibole forms of asbestos, chrysotile asbestos fail to accumulate in the lung tissue due to a mechanism involving fibers fragmentation in short pieces. Short exposure to chrysotile has not been associated with any histopathological alteration of lung tissue. Methods The present work focuses on the association of small chrysotile fibers with interphasic and mitotic human lung cancer cells in culture, using for analyses confocal laser scanning microscopy and 3D reconstructions. The main goal was to perform the analysis of abnormalities in mitosis of fibers-containing cells as well as to quantify nuclear DNA content of treated cells during their recovery in fiber-free culture medium. Results HK2 cells treated with chrysotile for 48 h and recovered in additional periods of 24, 48 and 72 h in normal medium showed increased frequency of multinucleated and apoptotic cells. DNA ploidy of the cells submitted to the same chrysotile treatment schedules showed enhanced aneuploidy values. The results were consistent with the high frequency of multipolar spindles observed and with the presence of fibers in the intercellular bridge during cytokinesis. Conclusion The present data show that 48 h chrysotile exposure can cause centrosome amplification, apoptosis and aneuploid cell formation even when long periods of recovery were provided. Internalized fibers seem to interact with the chromatin during mitosis, and they could also interfere in cytokinesis, leading to cytokinesis failure which forms aneuploid or multinucleated cells with centrosome amplification.

  2. A Simplified Method for Three-Dimensional (3-D Ovarian Tissue Culture Yielding Oocytes Competent to Produce Full-Term Offspring in Mice.

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    Carolyn M Higuchi

    Full Text Available In vitro growth of follicles is a promising technology to generate large quantities of competent oocytes from immature follicles and could expand the potential of assisted reproductive technologies (ART. Isolated follicle culture is currently the primary method used to develop and mature follicles in vitro. However, this procedure typically requires complicated, time-consuming procedures, as well as destruction of the normal ovarian microenvironment. Here we describe a simplified 3-D ovarian culture system that can be used to mature multilayered secondary follicles into antral follicles, generating developmentally competent oocytes in vitro. Ovaries recovered from mice at 14 days of age were cut into 8 pieces and placed onto a thick Matrigel drop (3-D culture for 10 days of culture. As a control, ovarian pieces were cultured on a membrane filter without any Matrigel drop (Membrane culture. We also evaluated the effect of activin A treatment on follicle growth within the ovarian pieces with or without Matrigel support. Thus we tested four different culture conditions: C (Membrane/activin-, A (Membrane/activin+, M (Matrigel/activin-, and M+A (Matrigel/activin+. We found that the cultured follicles and oocytes steadily increased in size regardless of the culture condition used. However, antral cavity formation occurred only in the follicles grown in the 3-D culture system (M, M+A. Following ovarian tissue culture, full-grown GV oocytes were isolated from the larger follicles to evaluate their developmental competence by subjecting them to in vitro maturation (IVM and in vitro fertilization (IVF. Maturation and fertilization rates were higher using oocytes grown in 3-D culture (M, M+A than with those grown in membrane culture (C, A. In particular, activin A treatment further improved 3-D culture (M+A success. Following IVF, two-cell embryos were transferred to recipients to generate full-term offspring. In summary, this simple and easy 3-D ovarian

  3. Decrease of reactive oxygen species-related biomarkers in the tissue-mimic 3D spheroid culture of human lung cells exposed to zinc oxide nanoparticles.

    Science.gov (United States)

    Kim, Eunjoo; Jeon, Won Bae; Kim, Soonhyun; Lee, Soo-Keun

    2014-05-01

    Common 2-dimensional (2D) cell cultures do not adequately represent cell-cell and cell-matrix signaling and substantially different diffusion/transport pathways. To obtain tissue-mimic information on nanoparticle toxicity from in vitro cell tests, we used a 3-dimensional (3D) culture of human lung cells (A549) prepared with elastin-like peptides modified with an arginine-glycine-aspartate motif. The 3D cells showed different cellular phenotypes, gene expression profiles, and functionalities compared to the 2D cultured cells. In gene array analysis, 3D cells displayed the induced extracellular matrix (ECM)-related biological functions such as cell-to-cell signaling and interaction, cellular function and maintenance, connective tissue development and function, molecular transport, and tissue morphology. Additionally, the expression of ECM-related molecules, such as laminin, fibronectin, and insulin-like growth factor binding protein 3 (IGFBP3), was simultaneously induced at both mRNA and protein levels. When 0.08-50 microg/ml zinc oxide nanoparticles (ZnO-NPs) were administered to 2D and 3D cells, the cell proliferation was not significantly changed. The level of molecular markers for oxidative stress, such as superoxide dismutase (SOD), Bcl-2, ATP synthase, and Complex IV (cytochrome C oxidase), was significantly reduced in 2D culture when exposed to 10 microg/ml ZnO-NPs, but no significant decrease was detected in 3D culture when exposed to the same concentration of ZnO-NPs. In conclusion, the tissue-mimic phenotype and functionality of 3D cells could be achieved through the elevated expression of ECM components. The 3D cells were expected to help to better predict the nanotoxicity of ZnO-NPs at tissue-level by increased cell-cell and cell-ECM adhesion and signaling. The tissue-mimic morphology would also be useful to simulate the diffusion/transport of the nanoparticles in vitro.

  4. 3D co-cultures of keratinocytes and melanocytes and cytoprotective effects on keratinocytes against reactive oxygen species by insect virus-derived protein microcrystals.

    Science.gov (United States)

    Shimabukuro, Junji; Yamaoka, Ayako; Murata, Ken-Ichi; Kotani, Eiji; Hirano, Tomoko; Nakajima, Yumiko; Matsumoto, Goichi; Mori, Hajime

    2014-09-01

    Stable protein microcrystals called polyhedra are produced by certain insect viruses. Cytokines, such as fibroblast growth factors (FGFs), can be immobilized within polyhedra. Here, we investigated three-dimensional (3D) co-cultures of keratinocytes and melanocytes on collagen gel containing FGF-2 and FGF-7 polyhedra. Melanocytes were observed to reside at the base of the 3D cell culture and melanin was also typically observed in the lower layer. The 3D cell culture model with FGF-2 and FGF-7 polyhedra was a useful in vitro model of the epidermis due to effective melanogenesis, proliferation and differentiation of keratinocytes. FGF-7 polyhedra showed a potent cytoprotective effect when keratinocytes were treated with menadione, which is a generator of reactive oxygen species. The cytoprotective effect was activated by the inositol triphosphate kinase-Akt pathway leading to upregulation of the antioxidant enzymes superoxide dismutase and peroxiredoxin 6.

  5. Cytotoxic responses of carnosic acid and doxorubicin on breast cancer cells in butterfly-shaped microchips in comparison to 2D and 3D culture.

    Science.gov (United States)

    Yildiz-Ozturk, Ece; Gulce-Iz, Sultan; Anil, Muge; Yesil-Celiktas, Ozlem

    2017-04-01

    Two dimensional (2D) cell culture systems lack the ability to mimic in vivo conditions resulting in limitations for preclinical cell-based drug and toxicity screening assays and modelling tumor biology. Alternatively, 3D cell culture systems mimic the specificity of native tissue with better physiological integrity. In this regard, microfluidic chips have gained wide applicability for in vitro 3D cancer cell studies. The aim of this research was to develop a 3D biomimetic model comprising culture of breast cancer cells in butterfly-shaped microchip to determine the cytotoxicity of carnosic acid and doxorubicin on both estrogen dependent (MCF-7) and independent (MDA-MB231) breast cancer cells along with healthy mammary epithelial cells (MCF-10A) in 2D, 3D Matrigel™ and butterfly-shaped microchip environment. According to the developed mimetic model, carnosic acid exhibited a higher cytotoxicity towards MDA-MB 231, while doxorubicin was more effective against MCF-7. Although the cell viabilities were higher in comparison to 2D and 3D cell culture systems, the responses of the investigated molecules were different in the microchips based on the molecular weight and structural complexity indicating the importance of biomimicry in a physiologically relevant matrix.

  6. AN APPLICATION FOR CULTURAL HERITAGE IN ERASMUS PLACEMENT. SURVEYS AND 3D CATALOGING ARCHAEOLOGICAL FINDS IN MÉRIDA (SPAIN

    Directory of Open Access Journals (Sweden)

    S. Barba

    2012-09-01

    Full Text Available Man has always had the need to live with his past, with its places and its artefacts. The reconstructions, the economical changes, the urbanization and its speculations have devastated whole cities, changed the faces of their historical centers, changed the relationship between the new and the old. Also the millenarian 'rest' of the archaeological findings, and therefore the respect towards those ancient civilizations, has been troubled. Our continent is rich in masterpieces that the modern man are not able to protect and pass on to the future, it is commonplace to observe that the modern ‘civilization’ has cemented and suffocated the ancient city of Pompeii, or even worse, failed to protected it. Walking in the archaeological area of Paestum it can be noticed how just sixty years ago, no one had the slightest concern of fencing the amphitheatre and the Roman forum, or entire houses and shops, to lay a carpet of tar or simple to build constructions completely inferior compared to those majestic Greek temples. The engineers and the architects should be held responsible for this as based on their scientific and humanistic sensibility; they should bring together the man with his surroundings in the complete respects of the historical heritage. The interest in ancient began to change nearly three decades ago since it was realized that the "Cultural Heritage" is a major tourist attraction and, if properly managed and used, it can be an economical cornerstone. Today, thanks to survey and the 3D graphics, which provide powerful new tools, we are witnessing a new and real need for the conservation, cataloguing and enhancement as a way to revive our archaeological sites. As part of a major laboratory project, artefacts from the Roman period (I and II century b.C., found in the Spanish city of Mérida, declared World Heritage by UNESCO in 1993, were acquired with a 3D laser scanner VIVID 910, and then catalogued. Based on these brief comments we

  7. An Application for Cultural Heritage in Erasmus Placement. Surveys and 3d Cataloging Archaeological Finds in MÉRIDA (spain)

    Science.gov (United States)

    Barba, S.; Fiorillo, F.; Ortiz Coder, P.; D'Auria, S.; De Feo, E.

    2011-09-01

    Man has always had the need to live with his past, with its places and its artefacts. The reconstructions, the economical changes, the urbanization and its speculations have devastated whole cities, changed the faces of their historical centers, changed the relationship between the new and the old. Also the millenarian 'rest' of the archaeological findings, and therefore the respect towards those ancient civilizations, has been troubled. Our continent is rich in masterpieces that the modern man are not able to protect and pass on to the future, it is commonplace to observe that the modern `civilization' has cemented and suffocated the ancient city of Pompeii, or even worse, failed to protected it. Walking in the archaeological area of Paestum it can be noticed how just sixty years ago, no one had the slightest concern of fencing the amphitheatre and the Roman forum, or entire houses and shops, to lay a carpet of tar or simple to build constructions completely inferior compared to those majestic Greek temples. The engineers and the architects should be held responsible for this as based on their scientific and humanistic sensibility; they should bring together the man with his surroundings in the complete respects of the historical heritage. The interest in ancient began to change nearly three decades ago since it was realized that the "Cultural Heritage" is a major tourist attraction and, if properly managed and used, it can be an economical cornerstone. Today, thanks to survey and the 3D graphics, which provide powerful new tools, we are witnessing a new and real need for the conservation, cataloguing and enhancement as a way to revive our archaeological sites. As part of a major laboratory project, artefacts from the Roman period (I and II century b.C.), found in the Spanish city of Mérida, declared World Heritage by UNESCO in 1993, were acquired with a 3D laser scanner VIVID 910, and then catalogued. Based on these brief comments we wanted to direct the work

  8. Leishmania amazonensis promastigotes in 3D Collagen I culture: an in vitro physiological environment for the study of extracellular matrix and host cell interactions

    Directory of Open Access Journals (Sweden)

    Debora B. Petropolis

    2014-04-01

    Full Text Available Leishmania amazonensis is the causative agent of American cutaneous leishmaniasis, an important neglected tropical disease. Once Leishmania amazonensis is inoculated into the human host, promastigotes are exposed to the extracellular matrix (ECM of the dermis. However, little is known about the interaction between the ECM and Leishmania promastigotes. In this study we established L. amazonensis promastigote culture in a three-dimensional (3D environment mainly composed of Collagen I (COL I. This 3D culture recreates in vitro some aspects of the human host infection site, enabling the study of the interaction mechanisms of L. amazonensis with the host ECM. Promastigotes exhibited “freeze and run” migration in the 3D COL I matrix, which is completely different from the conventional in vitro swimming mode of migration. Moreover, L. amazonensis promastigotes were able to invade, migrate inside, and remodel the 3D COL I matrix. Promastigote trans-matrix invasion and the freeze and run migration mode were also observed when macrophages were present in the matrix. At least two classes of proteases, metallo- and cysteine proteases, are involved in the 3D COL I matrix degradation caused by Leishmania. Treatment with a mixture of protease inhibitors significantly reduced promastigote invasion and migration through this matrix. Together our results demonstrate that L. amazonensis promastigotes release proteases and actively remodel their 3D environment, facilitating their migration. This raises the possibility that promastigotes actively interact with their 3D environment during the search for their cellular “home”—macrophages. Supporting this hypothesis, promastigotes migrated faster than macrophages in a novel 3D co-culture model.

  9. Measuring the impact of a 3D simulation experience on nursing students' cultural empathy using a modified version of the Kiersma-Chen Empathy Scale.

    Science.gov (United States)

    Everson, Naleya; Levett-Jones, Tracy; Lapkin, Samuel; Pitt, Victoria; van der Riet, Pamela; Rossiter, Rachel; Jones, Donovan; Gilligan, Conor; Courtney-Pratt, Helen

    2015-10-01

    To determine the effect of immersive 3D cultural simulation on nursing students' empathy towards culturally and linguistically diverse patients. Accelerated globalisation has seen a significant increase in cultural diversity in most regions of the world over the past forty years. Clinical encounters that do not acknowledge cultural factors contribute to adverse patient outcomes and health care inequities for culturally and linguistically diverse people. Cultural empathy is an antecedent to cultural competence. Thus, appropriate educational strategies are needed to enhance nursing students' cultural empathy and the capacity to deliver culturally competent care. A one-group pretest, post-test design was used for this study. The simulation exposed students to an unfolding scene in a hospital ward of a developing county. A convenience sample of second-year undergraduate nursing students (n = 460) from a semi-metropolitan university in Australia were recruited for the study. Characteristics of the sample were summarised using descriptive statistics. T-tests were performed to analyse the differences between pre- and post simulation empathy scores using an eight item modified version of the Kiersma-Chen Empathy Scale. Students' empathy towards culturally and linguistically diverse patients significantly improved after exposure to the 3D simulation experience. The mean scores for the Perspective Taking and Valuing Affective Empathy subscales also increased significantly postsimulation. The immersive 3D simulation had a positive impact on nursing students' empathy levels in regards to culturally and linguistically diverse groups. Research with other cohorts and in other contexts is required to further explore the impact of this educational approach. Immersive cultural simulation experiences offer opportunities to enhance the cultural empathy of nursing students. This may in turn have a positive impact on their cultural competence and consequently the quality of care they

  10. Novel MAPK-dependent and -independent tubulogenes identified via microarray analysis of 3D-cultured Madin-Darby canine kidney cells.

    Science.gov (United States)

    Chacon-Heszele, Maria F; Zuo, Xiaofeng; Hellman, Nathan E; McKenna, Sarah; Choi, Soo Young; Huang, Liwei; Tobias, John W; Park, Kwon Moo; Lipschutz, Joshua H

    2014-05-01

    Cystogenesis and tubulogenesis are basic building blocks for many epithelial organs, including the kidney. Most researchers have used two-dimensional (2D) cell culture to investigate signaling pathways downstream of hepatocyte growth factor (HGF). We hypothesize that three-dimensional (3D) collagen-grown Madin-Darby canine kidney (MDCK) cells, which form cysts and then tubulate in response to HGF, are a much more in vivo-like system for the identification of novel tubulogenes. With the use of a canine microarray containing over 20,000 genes, 2,417 genes were identified as potential tubulogenes that were differentially regulated, exclusively in 3D-grown MDCK cells. Among these, 840 were dependent on MAPK signaling. Importantly, this work shows that many putative tubulogenes, previously identified via microarray analysis of 2D cultures, including by us, do not change in 3D culture and vice versa. The use of a 3D-culture system allowed for the identification of novel MAPK-dependent and -independent genes that regulate early renal tubulogenesis in vitro, e.g., matrix metalloproteinase 1 (MMP1). Knockdown of MMP1 led to defects in cystogenesis and tubulogenesis in 3D-grown MDCK cells, most likely due to problems establishing normal polarity. We suggest that data obtained from 2D cultures, even those using MDCK cells treated with HGF, should not be automatically extrapolated to factors important for cystogenesis and tubulogenesis. Instead, 3D culture, which more closely replicates the biological environment and is therefore a more accurate model for identifying tubulogenes, is preferred. Results from the present analysis will be used to build a more accurate model of the signaling pathways that control cystogenesis and tubulogenesis.

  11. Multitemporal 3d Modelling for Cultural Heritage Emergency during Seismic Events: Damage Assesment of S. Agostino Church in Amatrice (ri)

    Science.gov (United States)

    Chiabrando, F.; Di Lolli, A.; Patrucco, G.; Spanò, A.; Sammartano, G.; Teppati Losè, L.

    2017-05-01

    One of the challenging purposes that must be undertaken by applied geomatics, is the need of monitoring by documenting continuously over time the evolution of urban spaces. Nowadays, this is a subject of great interest and study, mainly in case of sudden emergency events that implicate urban areas and specific historical buildings of our heritage. The newest Geomatics technique solutions must enable the demands of damage documentation, risk assessment, management and data sharing as efficiently as possible, in relation to the danger condition, to the accessibility constraints of areas and to the tight deadlines needs. In August 24th 2016, the first earthquake hit the area of central Italy with a magnitude of 6.0; since then, the earth never stop shaking in a wide area in the middle of Italy. On 26th and 30th of October, two other big seismic events were recorded (magnitude 5.9 and 6.5) and the already damaged built heritage were struck again. Since the beginning of the emergency all the available resources (human and material) were deployed and the world of researchers is trying to furnish an effective contribute as well. Politecnico di Torino, in coordination with the national institutions, is deploying people, expertise and resources. The geomatics research group and the connected Disaster Recovery team (DIRECT - http://areeweb.polito.it/direct/) is part of this process and is working in deep contact and collaboration with the Remotely Piloted Aircraft Systems (RPAS) group of the Italian Firefighter. Starting from the first earthquake the late medieval religious complex of S. Agostino has been carefully monitored and detected, using a multi-perspective oblique imagery strategy with the aim to achieve 3D aerial and terrestrial models, in a multi-temporal perspective concerning three different time situation.

  12. MULTITEMPORAL 3D MODELLING FOR CULTURAL HERITAGE EMERGENCY DURING SEISMIC EVENTS: DAMAGE ASSESMENT OF S. AGOSTINO CHURCH IN AMATRICE (RI

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    F. Chiabrando

    2017-05-01

    Full Text Available One of the challenging purposes that must be undertaken by applied geomatics, is the need of monitoring by documenting continuously over time the evolution of urban spaces. Nowadays, this is a subject of great interest and study, mainly in case of sudden emergency events that implicate urban areas and specific historical buildings of our heritage. The newest Geomatics technique solutions must enable the demands of damage documentation, risk assessment, management and data sharing as efficiently as possible, in relation to the danger condition, to the accessibility constraints of areas and to the tight deadlines needs. In August 24th 2016, the first earthquake hit the area of central Italy with a magnitude of 6.0; since then, the earth never stop shaking in a wide area in the middle of Italy. On 26th and 30th of October, two other big seismic events were recorded (magnitude 5.9 and 6.5 and the already damaged built heritage were struck again. Since the beginning of the emergency all the available resources (human and material were deployed and the world of researchers is trying to furnish an effective contribute as well. Politecnico di Torino, in coordination with the national institutions, is deploying people, expertise and resources. The geomatics research group and the connected Disaster Recovery team (DIRECT - http://areeweb.polito.it/direct/ is part of this process and is working in deep contact and collaboration with the Remotely Piloted Aircraft Systems (RPAS group of the Italian Firefighter. Starting from the first earthquake the late medieval religious complex of S. Agostino has been carefully monitored and detected, using a multi-perspective oblique imagery strategy with the aim to achieve 3D aerial and terrestrial models, in a multi-temporal perspective concerning three different time situation.

  13. Antitumor activity of amidino-substituted benzimidazole and benzimidazo[1,2-a]quinoline derivatives tested in 2D and 3D cell culture systems.

    Science.gov (United States)

    Brajša, Karmen; Vujasinović, Ines; Jelić, Dubravko; Trzun, Marija; Zlatar, Ivo; Karminski-Zamola, Grace; Hranjec, Marijana

    2016-12-01

    Due to a poor clinical predictive power of 2D cell cultures, standard tool for in vitro assays in drug discovery process, there is increasing interest in developing 3D in vitro cell cultures, biologically relevant assay feasible for the development of robust preclinical anti-cancer drug screening platforms. Herein, we tested amidino-substituted benzimidazoles and benzimidazo[1,2-a]quinolines as a small platform for comparison of antitumor activity in 2D and 3D cell culture systems and correlation with structure-activity relationship. 3D cell culture method was applied on a human cancer breast (SK-BR-3, MDA-MB-231, T-47D) and pancreatic cancer cells (MIA PaCa-2, PANC-1). Results obtained in 2D and 3D models were highly comparable, but in some cases we have observed significant disagreement indicating that some prominent compounds can be discarded in early phase of researching because of compounds with false positive result. To confirm which of cell culture systems is more accurate, in vivo profiling is needed.

  14. Increased extracellular matrix density decreases MCF10A breast cell acinus formation in 3D culture conditions.

    Science.gov (United States)

    Lance, Amanda; Yang, Chih-Chao; Swamydas, Muthulekha; Dean, Delphine; Deitch, Sandy; Burg, Karen J L; Dréau, Didier

    2016-01-01

    The extracellular matrix (ECM) contributes to the generation and dynamic of normal breast tissue, in particular to the generation of polarized acinar and ductal structures. In vitro 3D culture conditions, including variations in the composition of the ECM, have been shown to directly influence the formation and organization of acinus-like and duct-like structures. Furthermore, the density of the ECM appears to also play a role in the normal mammary tissue and tumour formation. Here we show that the density of the ECM directly influences the number, organization and function of breast acini. Briefly, non-malignant human breast MCF10A cells were incubated in increasing densities of a Matrigel®-collagen I matrix. Elastic moduli near and distant to the acinus structures were measured by atomic force microscopy, and the number of acinus structures was determined. Immunochemistry was used to investigate the expression levels of E-cadherin, laminin, matrix metalloproteinase-14 and ß-casein in MCF10A cells. The modulus of the ECM was significantly increased near the acinus structures and the number of acinus structures decreased with the increase in Matrigel-collagen I density. As evaluated by the expression of laminin, the organization of the acinus structures present was altered as the density of the ECM increased. Increases in both E-cadherin and MMP14 expression by MCF10A cells as ECM density increased were also observed. In contrast, MCF10A cells expressed lower ß-casein levels as the ECM density increased. Taken together, these observations highlight the key role of ECM density in modulating the number, organization and function of breast acini.

  15. The development of 3-D, in vitro, endothelial culture models for the study of coronary artery disease

    Directory of Open Access Journals (Sweden)

    Fraser Richard

    2009-10-01

    Full Text Available Abstract The response of the vascular endothelium to wall shear stress plays a central role in the development and progression of atherosclerosis. Current studies have investigated endothelial response using idealized in vitro flow chambers. Such cell culture models are unable to accurately replicate the complex in vivo wall shear stress patterns arising from anatomical geometries. To better understand this implication, we have created both simplified/tubular and anatomically realistic in vitro endothelial flow models of the human right coronary artery. A post-mortem vascular cast of the human left ventricular outflow tract was used to create geometrically accurate silicone elastomer models. Straight, tubular models were created using a custom made mold. Following the culture of human abdominal aortic endothelial cells within the inner lumen, cells were exposed to steady flow (Re = 233 for varying time periods. The resulting cell morphology was analyzed in terms of shape index and angle of orientation relative to the flow direction. In both models a progressive elongation and alignment of the endothelium in the flow direction was observed following 8, 12, and 24 hours. This change, however, was significantly less pronounced in the anatomical model (as observed from morphological variations indicative of localized flow features. Differences were also observed between the inner and outer walls at the disease-prone proximal region. Since morphological adaptation is a visual indication of endothelial shear stress activation, the use of anatomical models in endothelial genetic and biochemical studies may offer better insight into the disease process.

  16. Genotoxic Effects of Low- and High-LET Radiation on Human Epithelial Cells Grown in 2-D Versus 3-D Culture

    Science.gov (United States)

    Patel, Z. S.; Cucinotta, F. A.; Huff, J. L.

    2011-01-01

    Risk estimation for radiation-induced cancer relies heavily on human epidemiology data obtained from terrestrial irradiation incidents from sources such as medical and occupational exposures as well as from the atomic bomb survivors. No such data exists for exposures to the types and doses of high-LET radiation that will be encountered during space travel; therefore, risk assessment for space radiation requires the use of data derived from cell culture and animal models. The use of experimental models that most accurately replicate the response of human tissues is critical for precision in risk projections. This work compares the genotoxic effects of radiation on normal human epithelial cells grown in standard 2-D monolayer culture compared to 3-D organotypic co-culture conditions. These 3-D organotypic models mimic the morphological features, differentiation markers, and growth characteristics of fully-differentiated normal human tissue and are reproducible using defined components. Cultures were irradiated with 2 Gy low-LET gamma rays or varying doses of high-LET particle radiation and genotoxic damage was measured using a modified cytokinesis block micronucleus assay. Our results revealed a 2-fold increase in residual damage in 2 Gy gamma irradiated cells grown under organotypic culture conditions compared to monolayer culture. Irradiation with high-LET particle radiation gave similar results, while background levels of damage were comparable under both scenarios. These observations may be related to the phenomenon of "multicellular resistance" where cancer cells grown as 3-D spheroids or in vivo exhibit an increased resistance to killing by chemotherapeutic agents compared to the same cells grown in 2-D culture. A variety of factors are likely involved in mediating this process, including increased cell-cell communication, microenvironment influences, and changes in cell cycle kinetics that may promote survival of damaged cells in 3-D culture that would

  17. State-of-The-Art and Applications of 3D Imaging Sensors in Industry, Cultural Heritage, Medicine, and Criminal Investigation

    Science.gov (United States)

    Sansoni, Giovanna; Trebeschi, Marco; Docchio, Franco

    2009-01-01

    3D imaging sensors for the acquisition of three dimensional (3D) shapes have created, in recent years, a considerable degree of interest for a number of applications. The miniaturization and integration of the optical and electronic components used to build them have played a crucial role in the achievement of compactness, robustness and flexibility of the sensors. Today, several 3D sensors are available on the market, even in combination with other sensors in a “sensor fusion” approach. An importance equal to that of physical miniaturization has the portability of the measurements, via suitable interfaces, into software environments designed for their elaboration, e.g., CAD-CAM systems, virtual renders, and rapid prototyping tools. In this paper, following an overview of the state-of-art of 3D imaging sensors, a number of significant examples of their use are presented, with particular reference to industry, heritage, medicine, and criminal investigation applications. PMID:22389618

  18. Application of standard cell cultures and 3D in vitro tissue models as an effective tool in drug design and development.

    Science.gov (United States)

    Amelian, Aleksandra; Wasilewska, Katarzyna; Megias, Diego; Winnicka, Katarzyna

    2017-03-23

    Cell culture systems are essential tools used in a wide range of biomedical and clinical studies. Two dimensional cell culture models (2D) provide basic information on cytotoxicity, penetration and accumulation of drugs in cells and they are of outmost importance when selecting new compounds of the desired biopharmaceutical properties as candidates for novel drugs. The improvement over 2D growing cells are three dimensional (3D) tissue models that mimic in vivo conditions and the functions of living tissue more accurately. These models reduce the cost of drug development, enable more efficient drug screening, minimise failure rate in medicine discovery and eliminate animal use during experiments. The article provides an overview of 2D cell cultures and 3D tissue models - their properties, basic procedures, conditions of culturing and applications. Copyright © 2017 Institute of Pharmacology, Polish Academy of Sciences. Published by Elsevier Urban & Partner Sp. z o.o. All rights reserved.

  19. PCaAnalyser: a 2D-image analysis based module for effective determination of prostate cancer progression in 3D culture.

    Directory of Open Access Journals (Sweden)

    Md Tamjidul Hoque

    Full Text Available Three-dimensional (3D in vitro cell based assays for Prostate Cancer (PCa research are rapidly becoming the preferred alternative to that of conventional 2D monolayer cultures. 3D assays more precisely mimic the microenvironment found in vivo, and thus are ideally suited to evaluate compounds and their suitability for progression in the drug discovery pipeline. To achieve the desired high throughput needed for most screening programs, automated quantification of 3D cultures is required. Towards this end, this paper reports on the development of a prototype analysis module for an automated high-content-analysis (HCA system, which allows for accurate and fast investigation of in vitro 3D cell culture models for PCa. The Java based program, which we have named PCaAnalyser, uses novel algorithms that allow accurate and rapid quantitation of protein expression in 3D cell culture. As currently configured, the PCaAnalyser can quantify a range of biological parameters including: nuclei-count, nuclei-spheroid membership prediction, various function based classification of peripheral and non-peripheral areas to measure expression of biomarkers and protein constituents known to be associated with PCa progression, as well as defining segregate cellular-objects effectively for a range of signal-to-noise ratios. In addition, PCaAnalyser architecture is highly flexible, operating as a single independent analysis, as well as in batch mode; essential for High-Throughput-Screening (HTS. Utilising the PCaAnalyser, accurate and rapid analysis in an automated high throughput manner is provided, and reproducible analysis of the distribution and intensity of well-established markers associated with PCa progression in a range of metastatic PCa cell-lines (DU145 and PC3 in a 3D model demonstrated.

  20. 3D co-cultures of keratinocytes and melanocytes and cytoprotective effects on keratinocytes against reactive oxygen species by insect virus-derived protein microcrystals

    Energy Technology Data Exchange (ETDEWEB)

    Shimabukuro, Junji; Yamaoka, Ayako; Murata, Ken-ichi [Department of Applied Biology, Kyoto Institute of Technology, Kyoto (Japan); Kotani, Eiji [Department of Applied Biology, Kyoto Institute of Technology, Kyoto (Japan); Insect Biomedical Research Center, Kyoto Institute of Technology, Kyoto (Japan); Hirano, Tomoko [Venture Laboratory, Kyoto Institute of Technology, Kyoto (Japan); Nakajima, Yumiko [Functional Genomics Group, COMB, Tropical Biosphere Research Center, University of the Ryukyus, Okinawa (Japan); Matsumoto, Goichi [Division of Oral Surgery, Yokohama Clinical Education Center of Kanagawa Dental University, Yokohama (Japan); Mori, Hajime, E-mail: hmori@kit.ac.jp [Department of Applied Biology, Kyoto Institute of Technology, Kyoto (Japan); Insect Biomedical Research Center, Kyoto Institute of Technology, Kyoto (Japan)

    2014-09-01

    Stable protein microcrystals called polyhedra are produced by certain insect viruses. Cytokines, such as fibroblast growth factors (FGFs), can be immobilized within polyhedra. Here, we investigated three-dimensional (3D) co-cultures of keratinocytes and melanocytes on collagen gel containing FGF-2 and FGF-7 polyhedra. Melanocytes were observed to reside at the base of the 3D cell culture and melanin was also typically observed in the lower layer. The 3D cell culture model with FGF-2 and FGF-7 polyhedra was a useful in vitro model of the epidermis due to effective melanogenesis, proliferation and differentiation of keratinocytes. FGF-7 polyhedra showed a potent cytoprotective effect when keratinocytes were treated with menadione, which is a generator of reactive oxygen species. The cytoprotective effect was activated by the inositol triphosphate kinase–Akt pathway leading to upregulation of the antioxidant enzymes superoxide dismutase and peroxiredoxin 6. - Highlights: • 3D cultures using FGF-2 and FGF-7 microcrystals as a human skin model • Cytoprotection of keratinocytes against ROS by FGF-7 microcrystals • Overexpression of SOD and Prdx6 in keratinocytes by FGF-7 microcrystals.

  1. Regulation of adipose-tissue-derived stromal cell orientation and motility in 2D- and 3D-cultures by direct-current electrical field.

    Science.gov (United States)

    Yang, Gang; Long, Haiyan; Ren, Xiaomei; Ma, Kunlong; Xiao, Zhenghua; Wang, Ying; Guo, Yingqiang

    2017-02-01

    Cell alignment and motility play a critical role in a variety of cell behaviors, including cytoskeleton reorganization, membrane-protein relocation, nuclear gene expression, and extracellular matrix remodeling. Direct current electric field (EF) in vitro can direct many types of cells to align vertically to EF vector. In this work, we investigated the effects of EF stimulation on rat adipose-tissue-derived stromal cells (ADSCs) in 2D-culture on plastic culture dishes and in 3D-culture on various scaffold materials, including collagen hydrogels, chitosan hydrogels and poly(L-lactic acid)/gelatin electrospinning fibers. Rat ADSCs were exposed to various physiological-strength EFs in a homemade EF-bioreactor. Changes of morphology and movements of cells affected by applied EFs were evaluated by time-lapse microphotography, and cell survival rates and intracellular calcium oscillations were also detected. Results showed that EF facilitated ADSC morphological changes, under 6 V/cm EF strength, and that ADSCs in 2D-culture aligned vertically to EF vector and kept a good cell survival rate. In 3D-culture, cell galvanotaxis responses were subject to the synergistic effect of applied EF and scaffold materials. Fast cell movement and intracellular calcium activities were observed in the cells of 3D-culture. We believe our research will provide some experimental references for the future study in cell galvanotaxis behaviors.

  2. A novel 3D fibril force assay implicates src in tumor cell force generation in collagen networks.

    Directory of Open Access Journals (Sweden)

    Robert J Polackwich

    Full Text Available New insight into the biomechanics of cancer cell motility in 3D extracellular matrix (ECM environments would significantly enhance our understanding of aggressive cancers and help identify new targets for intervention. While several methods for measuring the forces involved in cell-matrix interactions have been developed, previous to this study none have been able to measure forces in a fibrillar environment. We have developed a novel assay for simultaneously measuring cell mechanotransduction and motility in 3D fibrillar environments. The assay consists of a controlled-density fibrillar collagen gel atop a controlled-stiffness polyacrylamide (PAA surface. Forces generated by living cells and their migration in the 3D collagen gel were measured with the 3D motion of tracer beads within the PAA layer. Here, this 3D fibril force assay is used to study the role of the invasion-associated protein kinase Src in mechanotransduction and motility. Src expression and activation are linked with proliferation, invasion, and metastasis, and have been shown to be required in 2D for invadopodia membranes to direct and mediate invasion. Breast cancer cell line MDA-MD-231 was stably transfected with GFP-tagged constitutively active Src or wild-type Src. In 3D fibrillar collagen matrices we found that, relative to wild-type Src, constitutively active Src: 1 increased the strength of cell-induced forces on the ECM, 2 did not significantly change migration speed, and 3 increased both the duration and the length, but not the number, of long membrane protrusions. Taken together, these results support the hypothesis that Src controls invasion by controlling the ability of the cell to form long lasting cellular protrusions to enable penetration through tissue barriers, in addition to its role in promoting invadopodia matrix-degrading activity.

  3. Modeling the Decay in AN Hbim Starting from 3d Point Clouds. a Followed Approach for Cultural Heritage Knowledge

    Science.gov (United States)

    Chiabrando, F.; Lo Turco, M.; Rinaudo, F.

    2017-08-01

    The recent trends in architectural data management imply the scientific and professional collaborations of several disciplines involved in the design, restoration and maintenance. It seems an achieved concept that, in the next future, all the information connected to new interventions or conservation activities on historical buildings will be managed by using a BIM platform. Nowadays the actual range or image based metric survey techniques (mainly produced by using Terrestrial Laser Scanner or photogrammetric platform today more based on projective geometry) allow to generate 3D point clouds, 3D models, orthophotos and other outputs with assessed accuracy. The subsequent conversion of 3D information into parametric components, especially in an historical environment, is not easy and has a lot of open issues. According to the actual BIM commercial software and to the embedded tools or plugin, the paper deals with the methodology followed for the realization of two parametric 3D models (Palazzo Sarmatoris and Smistamento RoundHouse, two historical building in the north-west part of Italy). The paper describes the proposed workflow according to the employed plug-in for automatic reconstruction and to the solution adopted for the well-known problems connected to the modeling phase such as the vaults realization or the 3D irregular surfaces modeling. Finally, the studied strategy for mapping the decay in a BIM environment and the connected results with the conclusions and future perspectives are critically discussed.

  4. MODELING THE DECAY IN AN HBIM STARTING FROM 3D POINT CLOUDS. A FOLLOWED APPROACH FOR CULTURAL HERITAGE KNOWLEDGE

    Directory of Open Access Journals (Sweden)

    F. Chiabrando

    2017-08-01

    Full Text Available The recent trends in architectural data management imply the scientific and professional collaborations of several disciplines involved in the design, restoration and maintenance. It seems an achieved concept that, in the next future, all the information connected to new interventions or conservation activities on historical buildings will be managed by using a BIM platform. Nowadays the actual range or image based metric survey techniques (mainly produced by using Terrestrial Laser Scanner or photogrammetric platform today more based on projective geometry allow to generate 3D point clouds, 3D models, orthophotos and other outputs with assessed accuracy. The subsequent conversion of 3D information into parametric components, especially in an historical environment, is not easy and has a lot of open issues. According to the actual BIM commercial software and to the embedded tools or plugin, the paper deals with the methodology followed for the realization of two parametric 3D models (Palazzo Sarmatoris and Smistamento RoundHouse, two historical building in the north-west part of Italy. The paper describes the proposed workflow according to the employed plug-in for automatic reconstruction and to the solution adopted for the well-known problems connected to the modeling phase such as the vaults realization or the 3D irregular surfaces modeling. Finally, the studied strategy for mapping the decay in a BIM environment and the connected results with the conclusions and future perspectives are critically discussed.

  5. Facial recognition software success rates for the identification of 3D surface reconstructed facial images: implications for patient privacy and security.

    Science.gov (United States)

    Mazura, Jan C; Juluru, Krishna; Chen, Joseph J; Morgan, Tara A; John, Majnu; Siegel, Eliot L

    2012-06-01

    Image de-identification has focused on the removal of textual protected health information (PHI). Surface reconstructions of the face have the potential to reveal a subject's identity even when textual PHI is absent. This study assessed the ability of a computer application to match research subjects' 3D facial reconstructions with conventional photographs of their face. In a prospective study, 29 subjects underwent CT scans of the head and had frontal digital photographs of their face taken. Facial reconstructions of each CT dataset were generated on a 3D workstation. In phase 1, photographs of the 29 subjects undergoing CT scans were added to a digital directory and tested for recognition using facial recognition software. In phases 2-4, additional photographs were added in groups of 50 to increase the pool of possible matches and the test for recognition was repeated. As an internal control, photographs of all subjects were tested for recognition against an identical photograph. Of 3D reconstructions, 27.5% were matched correctly to corresponding photographs (95% upper CL, 40.1%). All study subject photographs were matched correctly to identical photographs (95% lower CL, 88.6%). Of 3D reconstructions, 96.6% were recognized simply as a face by the software (95% lower CL, 83.5%). Facial recognition software has the potential to recognize features on 3D CT surface reconstructions and match these with photographs, with implications for PHI.

  6. The implications of free 3D scanning in the conservation state assessment of old wood painted icon

    Science.gov (United States)

    Munteanu, Marius; Sandu, Ion

    2016-06-01

    The present paper presents the conservation state and the making of a 3D model of a XVIII-th century orthodox icon on wood support, using free available software and cloud computing. In order to create the 3D model of the painting layer of the icon a number of 70 pictures were taken using a Nikon DSLR D3300, 24.2 MP in setup with a Hama Star 75 photo tripod, in loops 360° around the painting, at three different angles. The pictures were processed with Autodesk I23D Catch, which automatically finds and matches common features among all of the uploaded photographs in order to create the 3D scene, using the power and speed of cloud computing. The obtained 3D model was afterwards analyzed and processed in order to obtain a final version, which can now be use to better identify, to map and to prioritize the future conservation processes and finally can be shared online as an animation.

  7. Differential effects of MAPK pathway inhibitors on migration and invasiveness of BRAF(V600E) mutant thyroid cancer cells in 2D and 3D culture.

    Science.gov (United States)

    Ingeson-Carlsson, Camilla; Martinez-Monleon, Angela; Nilsson, Mikael

    2015-11-01

    Tumor microenvironment influences targeted drug therapy. In this study we compared drug responses to RAF and MEK inhibitors on tumor cell migration in 2D and 3D culture of BRAF(V600E) mutant cell lines derived from human papillary (BCPAP) and anaplastic (SW1736) thyroid carcinomas. Scratch wounding was compared to a double-layered collagen gel model developed for analysis of directed tumor cell invasion during prolonged culture. In BCPAP both PLX4720 and U0126 inhibited growth and migration in 2D and decreased tumor cell survival in 3D. In SW1736 drugs had no effect on migration in 2D but decreased invasion in 3D, however this related to reduced growth. Dual inhibition of BRAF(V600E) and MEK reduced but did not prevent SW1736 invasion although rebound phosphorylation of ERK in response to PLX4720 was blocked by U0126. These findings indicate that anti-tumor drug effects in vitro differ depending on culture conditions (2D vs. 3D) and that the invasive features of anaplastic thyroid cancer depend on non-MEK mechanism(s).

  8. Radiochromic 3D Detectors

    Science.gov (United States)

    Oldham, Mark

    2015-01-01

    Radiochromic materials exhibit a colour change when exposed to ionising radiation. Radiochromic film has been used for clinical dosimetry for many years and increasingly so recently, as films of higher sensitivities have become available. The two principle advantages of radiochromic dosimetry include greater tissue equivalence (radiologically) and the lack of requirement for development of the colour change. In a radiochromic material, the colour change arises direct from ionising interactions affecting dye molecules, without requiring any latent chemical, optical or thermal development, with important implications for increased accuracy and convenience. It is only relatively recently however, that 3D radiochromic dosimetry has become possible. In this article we review recent developments and the current state-of-the-art of 3D radiochromic dosimetry, and the potential for a more comprehensive solution for the verification of complex radiation therapy treatments, and 3D dose measurement in general.

  9. Novel 3D Culture Systems for Studies of Human Liver Function and Assessments of the Hepatotoxicity of Drugs and Drug Candidates.

    Science.gov (United States)

    Lauschke, Volker M; Hendriks, Delilah F G; Bell, Catherine C; Andersson, Tommy B; Ingelman-Sundberg, Magnus

    2016-12-19

    The liver is an organ with critical importance for drug treatment as the disposition and response to a given drug is often determined by its hepatic metabolism. Patient-specific factors can entail increased susceptibility to drug-induced liver injury, which constitutes a major risk for drug development programs causing attrition of promising drug candidates or costly withdrawals in postmarketing stages. Hitherto, mainly animal studies and 2D hepatocyte systems have been used for the examination of human drug metabolism and toxicity. Yet, these models are far from satisfactory due to extensive species differences and because hepatocytes in 2D cultures rapidly dedifferentiate resulting in the loss of their hepatic phenotype and functionality. With the increasing comprehension that 3D cell culture systems more accurately reflect in vivo physiology, in the recent decade more and more research has focused on the development and optimization of various 3D culture strategies in an attempt to preserve liver properties in vitro. In this contribution, we critically review these developments, which have resulted in an arsenal of different static and perfused 3D models. These systems include sandwich-cultured hepatocytes, spheroid culture platforms, and various microfluidic liver or multiorgan biochips. Importantly, in many of these models hepatocytes maintain their phenotype for prolonged times, which allows probing the potential of newly developed chemical entities to cause chronic hepatotoxicity. Moreover, some platforms permit the investigation of drug action in specific genetic backgrounds or diseased hepatocytes, thereby significantly expanding the repertoire of tools to detect drug-induced liver injuries. It is concluded that the development of 3D liver models has hitherto been fruitful and that systems are now at hand whose sensitivity and specificity in detecting hepatotoxicity are superior to those of classical 2D culture systems. For the future, we highlight the

  10. State-of-The-Art and Applications of 3D Imaging Sensors in Industry, Cultural Heritage, Medicine, and Criminal Investigation

    Directory of Open Access Journals (Sweden)

    Giovanna Sansoni

    2009-01-01

    Full Text Available 3D imaging sensors for the acquisition of three dimensional (3D shapes have created, in recent years, a considerable degree of interest for a number of applications. The miniaturization and integration of the optical and electronic components used to build them have played a crucial role in the achievement of compactness, robustness and flexibility of the sensors. Today, several 3D sensors are available on the market, even in combination with other sensors in a “sensor fusion” approach. An importance equal to that of physical miniaturization has the portability of the measurements, via suitable interfaces, into software environments designed for their elaboration, e.g., CAD-CAM systems, virtual renders, and rapid prototyping tools. In this paper, following an overview of the state-of-art of 3D imaging sensors, a number of significant examples of their use are presented, with particular reference to industry, heritage, medicine, and criminal investigation applications.

  11. 3DMADMAC|AUTOMATED: synergistic hardware and software solution for automated 3D digitization of cultural heritage objects

    Directory of Open Access Journals (Sweden)

    Robert Sitnik

    2011-12-01

    Full Text Available In this article a fully automated 3D shape measurement system and data processing algorithms are presented. Main purpose of this system is to automatically (without any user intervention and rapidly (at least ten times faster than manual measurement digitize whole object’s surface with some limitations to its properties: maximum measurement volume is described as a cylinder with 2,8m height and 0,6m radius, maximum object's weight is 2 tons.  Measurement head is automatically calibrated by the system for chosen working volume (from 120mm x 80mm x 60mm and ends up to 1,2m x 0,8m x 0,6m. Positioning of measurement head in relation to measured object is realized by computer-controlled manipulator. The system is equipped with two independent collision detection modules to prevent damaging measured object with moving sensor’s head. Measurement process is divided into three steps. First step is used for locating any part of object’s surface in assumed measurement volume. Second step is related to calculation of "next best view" position of measurement head on the base of existing 3D scans. Finally small holes in measured 3D surface are detected and measured. All 3D data processing (filtering, ICP based fitting and final views integration is performed automatically. Final 3D model is created on the base of user specified parameters like accuracy of surface representation and/or density of surface sampling. In the last section of the paper, exemplary measurement result of two objects: biscuit (from the collection of Museum Palace at Wilanów and Roman votive altar (Lower Moesia, II-III AD are presented.

  12. Generating 3D anatomically detailed models of the retina from OCT data sets: implications for computational modelling

    Science.gov (United States)

    Shalbaf, Farzaneh; Dokos, Socrates; Lovell, Nigel H.; Turuwhenua, Jason; Vaghefi, Ehsan

    2015-12-01

    Retinal prosthesis has been proposed to restore vision for those suffering from the retinal pathologies that mainly affect the photoreceptors layer but keep the inner retina intact. Prior to costly risky experimental studies computational modelling of the retina will help to optimize the device parameters and enhance the outcomes. Here, we developed an anatomically detailed computational model of the retina based on OCT data sets. The consecutive OCT images of individual were subsequently segmented to provide a 3D representation of retina in the form of finite elements. Thereafter, the electrical properties of the retina were modelled by implementing partial differential equation on the 3D mesh. Different electrode configurations, that is bipolar and hexapolar configurations, were implemented and the results were compared with the previous computational and experimental studies. Furthermore, the possible effects of the curvature of retinal layers on the current steering through the retina were proposed and linked to the clinical observations.

  13. A complex 3D human tissue culture system based on mammary stromal cells and silk scaffolds for modeling breast morphogenesis and function.

    Science.gov (United States)

    Wang, Xiuli; Sun, Lin; Maffini, Maricel V; Soto, Ana; Sonnenschein, Carlos; Kaplan, David L

    2010-05-01

    Epithelial-stromal interactions play a crucial role in normal embryonic development and carcinogenesis of the human breast while the underlying mechanisms of these events remain poorly understood. To address this issue, we constructed a physiologically relevant, three-dimensional (3D) culture surrogate of complex human breast tissue that included a tri-culture system made up of human mammary epithelial cells (MCF10A), human fibroblasts and adipocytes, i.e., the two dominant breast stromal cell types, in a Matrigel/collagen mixture on porous silk protein scaffolds. The presence of stromal cells inhibited MCF10A cell proliferation and induced both alveolar and ductal morphogenesis and enhanced casein expression. In contrast to the immature polarity exhibited by co-cultures with either fibroblasts or adipocytes, the alveolar structures formed by the tri-cultures exhibited proper polarity similar to that observed in breast tissue in vivo. Only alveolar structures with reverted polarity were observed in MCF10A monocultures. Consistent with their phenotypic appearance, more functional differentiation of epithelial cells was also observed in the tri-cultures, where casein alpha- and -beta mRNA expression was significantly increased. This in vitro tri-culture breast tissue system sustained on silk scaffold effectively represents a more physiologically relevant 3D microenvironment for mammary epithelial cells and stromal cells than either co-cultures or monocultures. This experimental model provides an important first step for bioengineering an informative human breast tissue system, with which to study normal breast morphogenesis and neoplastic transformation.

  14. Osteoinduction and survival of osteoblasts and bone-marrow stromal cells in 3D biphasic calcium phosphate scaffolds under static and dynamic culture conditions.

    Science.gov (United States)

    Rath, Subha N; Strobel, Leonie A; Arkudas, Andreas; Beier, Justus P; Maier, Anne-Kathrin; Greil, Peter; Horch, Raymund E; Kneser, Ulrich

    2012-10-01

    In many tissue engineering approaches, the basic difference between in vitro and in vivo conditions for cells within three-dimensional (3D) constructs is the nutrition flow dynamics. To achieve comparable results in vitro, bioreactors are advised for improved cell survival, as they are able to provide a controlled flow through the scaffold. We hypothesize that a bioreactor would enhance long-term differentiation conditions of osteogenic cells in 3D scaffolds. To achieve this either primary rat osteoblasts or bone marrow stromal cells (BMSC) were implanted on uniform-sized biphasic calcium phosphate (BCP) scaffolds produced by a 3D printing method. Three types of culture conditions were applied: static culture without osteoinduction (Group A); static culture with osteoinduction (Group B); dynamic culture with osteoinduction (Group C). After 3 and 6 weeks, the scaffolds were analysed by alkaline phosphatase (ALP), dsDNA amount, SEM, fluorescent labelled live-dead assay, and real-time RT-PCR in addition to weekly alamarBlue assays. With osteoinduction, increased ALP values and calcium deposition are observed; however, under static conditions, a significant decrease in the cell number on the biomaterial is observed. Interestingly, the bioreactor system not only reversed the decreased cell numbers but also increased their differentiation potential. We conclude from this study that a continuous flow bioreactor not only preserves the number of osteogenic cells but also keeps their differentiation ability in balance providing a suitable cell-seeded scaffold product for applications in regenerative medicine.

  15. Oxygen Partial Pressure Is a Rate-Limiting Parameter for Cell Proliferation in 3D Spheroids Grown in Physioxic Culture Condition.

    Science.gov (United States)

    Gomes, Aurélie; Guillaume, Ludivine; Grimes, David Robert; Fehrenbach, Jérôme; Lobjois, Valérie; Ducommun, Bernard

    2016-01-01

    The in situ oxygen partial pressure in normal and tumor tissues is in the range of a few percent. Therefore, when studying cell growth in 3D culture systems, it is essential to consider how the physiological oxygen concentration, rather than the one in the ambient air, influences the proliferation parameters. Here, we investigated the effect of reducing oxygen partial pressure from 21% to 5% on cell proliferation rate and regionalization in a 3D tumor spheroid model. We found that 5% oxygen concentration strongly inhibited spheroid growth, changed the proliferation gradient and reduced the 50% In Depth Proliferation index (IDP50), compared with culture at 21% oxygen. We then modeled the oxygen partial pressure profiles using the experimental data generated by culturing spheroids in physioxic and normoxic conditions. Although hypoxia occurred at similar depth in spheroids grown in the two conditions, oxygen partial pressure was a major rate-limiting factor with a critical effect on cell proliferation rate and regionalization only in spheroids grown in physioxic condition and not in spheroids grown at atmospheric normoxia. Our findings strengthen the need to consider conducting experiment in physioxic conditions (i.e., tissue normoxia) for proper understanding of cancer cell biology and the evaluation of anticancer drugs in 3D culture systems.

  16. Low-cost production and sealing procedure of mechanical parts of a versatile 3D-printed perfusion chamber for digital holographic microscopy of primary neurons in culture

    Science.gov (United States)

    Bélanger, Erik; Lévesque, Sébastien A.; Anctil, Gabriel; Poulin-Girard, Anne-Sophie; Marquet, Pierre

    2017-02-01

    We have developed a prototype of a low-cost and versatile 3D-printed perfusion chamber for digital holographic microscopy (DHM) of primary neurons in culture. The imaging chamber is 3D-printed in biocompatible plastic. It is easily convertible between a closed configuration, for refractive index - cellular thickness decoupling, and an open configuration, for electrophysiology. In the closed arrangement, the imaging volume is small, allowing a rapid laminar flow with a fast turnover for an optimal implementation of the decoupling procedure. This paper highlights especially the challenges faced while designing and prototyping the 3D-printed closed perfusion chamber with a small imaging volume for DHM. As all 3D-printed mechanical parts were initially leaking because of internal porosities, we developed a simple sealing protocol using acetone vapors to smooth surfaces. Using this protocol, almost all mechanical parts were successfully sealed. Therefore, the production process of the actual prototype, i.e. the 3D printing and the sealing method, is satisfactory for our target application in the field of microfluidics.

  17. Optical sensing technologies for the generation of reality-based 3D models of Cultural Heritage artifacts

    OpenAIRE

    2012-01-01

    The central theme of the thesis is the use of triangulation laser scanning and other optical three-dimensional surveying systems for the realization of 3D models of objects of historical and artistic interest - sculptures, archaeological finds, decorative elements in architecture. The subject is faced keeping in mind the purposes and needs the models should or could meet, and which are the challenging steps to become a more common tool. To name one, a fundamental aspect is bridging gaps betwe...

  18. Ground and Aerial Digital Documentation of Cultural Heritage: Providing Tools for 3d Exploitation of Archaeological Data

    Science.gov (United States)

    Cantoro, G.

    2017-02-01

    Archaeology is by its nature strictly connected with the physical landscape and as such it explores the inter-relations of individuals with places in which they leave and the nature that surrounds them. Since its earliest stages, archaeology demonstrated its permeability to scientific methods and innovative techniques or technologies. Archaeologists were indeed between the first to adopt GIS platforms (since already almost three decades) on large scale and are now between the most demanding customers for emerging technologies such as digital photogrammetry and drone-aided aerial photography. This paper aims at presenting case studies where the "3D approach" can be critically analysed and compared with more traditional means of documentation. Spot-light is directed towards the benefits of a specifically designed platform for user to access the 3D point-clouds and explore their characteristics. Beside simple measuring and editing tools, models are presented in their actual context and location, with historical and archaeological information provided on the side. As final step of a parallel project on geo-referencing and making available a large archive of aerial photographs, 3D models derived from photogrammetric processing of images have been uploaded and linked to photo-footprints polygons. Of great importance in such context is the possibility to interchange the point-cloud colours with satellite imagery from OpenLayers. This approach makes it possible to explore different landscape configurations due to time-changes with simple clicks. In these cases, photogrammetry or 3D laser scanning replaced, sided or integrated legacy documentation, creating at once a new set of information for forthcoming research and ideally new discoveries.

  19. 3D micro-XRF for cultural heritage objects: new analysis strategies for the investigation of the Dead Sea Scrolls.

    Science.gov (United States)

    Mantouvalou, Ioanna; Wolff, Timo; Hahn, Oliver; Rabin, Ira; Lühl, Lars; Pagels, Marcel; Malzer, Wolfgang; Kanngiesser, Birgit

    2011-08-15

    A combination of 3D micro X-ray fluorescence spectroscopy (3D micro-XRF) and micro-XRF was utilized for the investigation of a small collection of highly heterogeneous, partly degraded Dead Sea Scroll parchment samples from known excavation sites. The quantitative combination of the two techniques proves to be suitable for the identification of reliable marker elements which may be used for classification and provenance studies. With 3D micro-XRF, the three-dimensional nature, i.e. the depth-resolved elemental composition as well as density variations, of the samples was investigated and bromine could be identified as a suitable marker element. It is shown through a comparison of quantitative and semiquantitative values for the bromine content derived using both techniques that, for elements which are homogeneously distributed in the sample matrix, quantification with micro-XRF using a one-layer model is feasible. Thus, the possibility for routine provenance studies using portable micro-XRF instrumentation on a vast amount of samples, even on site, is obtained through this work.

  20. 3D Structural and Stratigraphic Architecture of the Northwest Santa Barbara Basin: Implications for Slope Stability and Submarine Landslide Occurrence

    Science.gov (United States)

    Wright, A.; Brothers, D. S.; Kluesner, J.; Johnson, S. Y.

    2016-12-01

    Multiple submarine landslides have been documented on the north flank of the Santa Barbara Basin and such failures are considered capable of generating local tsunami hazards to the Santa Barbara region. Past 2D seismic reflection data has provided a general view of the regional framework geology resulting from north-south compression, but fails to identify along-strike variations of faults and folds. This study uses industry 3D seismic reflection data encompassing the slope surrounding the 3.8 km2-Gaviota submarine landslide to investigate structural and stratigraphic controls of slope failure in this region. The 3D depth-migrated volume shows a complex network of faults that result in both broad and local zones of compression, folding, and uplift along the slope. One localized zone of enhanced anticlinal folding and uplift associated with small-scale thrust faults is located directly beneath the Gaviota slide, while another is beneath a seafloor fissure west of the slide inferred to represent incipient failure. New high-resolution 2D transects constrain the character of shallow deformation above the locally uplifted blocks. 3D isopach maps indicate the seafloor fissures trend along a key threshold of thickness between the seafloor and a shallow horizon; the fissures are also coincide with an apparent zone of shallow, gas-charged strata that onlap the steeply dipping flanks of local anticlinal deformation. Because the seafloor gradient near the Gaviota slide is significantly lower than the internal friction angle for fine-grained marine sediments, we propose that a combination of active deformation, sediment compaction, and gas charging acted to precondition the slope of the Gaviota landslide for failure by reducing the shear strength. Similar factors occur beneath intact sections of the slope adjacent to the slide, which should be considered prone to future landsliding.

  1. 3D DOCUMENTATION AND BIM MODELING OF CULTURAL HERITAGE STRUCTURES USING UAVS: THE CASE OF THE FOINIKARIA CHURCH

    OpenAIRE

    K. Themistocleous; A. Agapiou; D. Hadjimitsis

    2016-01-01

    The documentation of architectural cultural heritage sites has traditionally been expensive and labor-intensive. New innovative technologies, such as Unmanned Aerial Vehicles (UAVs), provide an affordable, reliable and straightforward method of capturing cultural heritage sites, thereby providing a more efficient and sustainable approach to documentation of cultural heritage structures. In this study, hundreds of images of the Panagia Chryseleousa church in Foinikaria, Cyprus were ta...

  2. 3D shape of asteroid (6) Hebe from VLT/SPHERE imaging: Implications for the origin of ordinary H chondrites

    Science.gov (United States)

    Marsset, M.; Carry, B.; Dumas, C.; Hanuš, J.; Viikinkoski, M.; Vernazza, P.; Müller, T. G.; Delbo, M.; Jehin, E.; Gillon, M.; Grice, J.; Yang, B.; Fusco, T.; Berthier, J.; Sonnett, S.; Kugel, F.; Caron, J.; Behrend, R.

    2017-08-01

    Context. The high-angular-resolution capability of the new-generation ground-based adaptive-optics camera SPHERE at ESO VLT allows us to assess, for the very first time, the cratering record of medium-sized (D 100-200 km) asteroids from the ground, opening the prospect of a new era of investigation of the asteroid belt's collisional history. Aims: We investigate here the collisional history of asteroid (6) Hebe and challenge the idea that Hebe may be the parent body of ordinary H chondrites, the most common type of meteorites found on Earth ( 34% of the falls). Methods: We observed Hebe with SPHERE as part of the science verification of the instrument. Combined with earlier adaptive-optics images and optical light curves, we model the spin and three-dimensional (3D) shape of Hebe and check the consistency of the derived model against available stellar occultations and thermal measurements. Results: Our 3D shape model fits the images with sub-pixel residuals and the light curves to 0.02 mag. The rotation period (7.274 47 h), spin (ECJ2000 λ, β of 343°, +47°), and volume-equivalent diameter (193 ± 6 km) are consistent with previous determinations and thermophysical modeling. Hebe's inferred density is 3.48 ± 0.64 g cm-3, in agreement with an intact interior based on its H-chondrite composition. Using the 3D shape model to derive the volume of the largest depression (likely impact crater), it appears that the latter is significantly smaller than the total volume of close-by S-type H-chondrite-like asteroid families. Conclusions: Our results imply that (6) Hebe is not the most likely source of H chondrites. Over the coming years, our team will collect similar high-precision shape measurements with VLT/SPHERE for 40 asteroids covering the main compositional classes, thus providing an unprecedented dataset to investigate the origin and collisional evolution of the asteroid belt. Based on observations made with ESO Telescopes at the La Silla Paranal Observatory

  3. Nonviral Gene Delivery of Growth and Differentiation Factor 5 to Human Mesenchymal Stem Cells Injected into a 3D Bovine Intervertebral Disc Organ Culture System

    Directory of Open Access Journals (Sweden)

    Christian Bucher

    2013-01-01

    Full Text Available Intervertebral disc (IVD cell therapy with unconditioned 2D expanded mesenchymal stem cells (MSC is a promising concept yet challenging to realize. Differentiation of MSCs by nonviral gene delivery of growth and differentiation factor 5 (GDF5 by electroporation mediated gene transfer could be an excellent source for cell transplantation. Human MSCs were harvested from bone marrow aspirate and GDF5 gene transfer was achieved by in vitro electroporation. Transfected cells were cultured as monolayers and as 3D cultures in 1.2% alginate bead culture. MSC expressed GDF5 efficiently for up to 21 days. The combination of GDF5 gene transfer and 3D culture in alginate showed an upregulation of aggrecan and SOX9, two markers for chondrogenesis, and KRT19 as a marker for discogenesis compared to untransfected cells. The cells encapsulated in alginate produced more proteoglycans expressed in GAG/DNA ratio. Furthermore, GDF5 transfected MCS injected into an IVD papain degeneration organ culture model showed a partial recovery of the GAG/DNA ratio after 7 days. In this study we demonstrate the potential of GDF5 transfected MSC as a promising approach for clinical translation for disc regeneration.

  4. Bound states in the 3d Ising model and implications for QCD at finite temperature and density

    CERN Document Server

    Caselle, M; Provero, P; Zarembo, K

    2002-01-01

    We study the spectrum of bound states of the three dimensional Ising model in the (h,beta) plane near the critical point. We show the existence of an unbinding line, defined as the boundary of the region where bound states exist. Numerical evidence suggests that this line coincides with the beta=beta_c axis. When the 3D Ising model is considered as an effective description of hot QCD at finite density, we conjecture the correspondence between the unbinding line and the line that separates the quark-gluon plasma phase from the superconducting phase. The bound states of the Ising model are conjectured to correspond to the diquarks of the latter phase of QCD.

  5. Drug-releasing nano-engineered titanium implants: therapeutic efficacy in 3D cell culture model, controlled release and stability.

    Science.gov (United States)

    Gulati, Karan; Kogawa, Masakazu; Prideaux, Matthew; Findlay, David M; Atkins, Gerald J; Losic, Dusan

    2016-12-01

    There is an ongoing demand for new approaches for treating localized bone pathologies. Here we propose a new strategy for treatment of such conditions, via local delivery of hormones/drugs to the trauma site using drug releasing nano-engineered implants. The proposed implants were prepared in the form of small Ti wires/needles with a nano-engineered oxide layer composed of array of titania nanotubes (TNTs). TNTs implants were inserted into a 3D collagen gel matrix containing human osteoblast-like, and the results confirmed cell migration onto the implants and their attachment and spread. To investigate therapeutic efficacy, TNTs/Ti wires loaded with parathyroid hormone (PTH), an approved anabolic therapeutic for the treatment of severe bone fractures, were inserted into 3D gels containing osteoblast-like cells. Gene expression studies revealed a suppression of SOST (sclerostin) and an increase in RANKL (receptor activator of nuclear factor kappa-B ligand) mRNA expression, confirming the release of PTH from TNTs at concentrations sufficient to alter cell function. The performance of the TNTs wire implants using an example of a drug needed at relatively higher concentrations, the anti-inflammatory drug indomethacin, is also demonstrated. Finally, the mechanical stability of the prepared implants was tested by their insertion into bovine trabecular bone cores ex vivo followed by retrieval, which confirmed the robustness of the TNT structures. This study provides proof of principle for the suitability of the TNT/Ti wire implants for localized bone therapy, which can be customized to cater for specific therapeutic requirements.

  6. CARS and SHG microscopy to follow the collagen production in living human corneal fibroblasts and mesenchymal stem cells in fibrin gel 3D cultures

    CERN Document Server

    Mortati, Leonardo; Sassi, Maria Paola

    2011-01-01

    Coherent anti-Stokes Raman scattering (CARS) microscopy is combined with second harmonic generation (SHG) technique in order to follow the early stage of stem cell differentiation within a 3D scaffold. CARS microscopy can detect lipid membranes and droplet compartments in living cells and SHG microscopy enables a strong imaging contrast for molecules with a non-centrosymmetric ordered structure like collagen. One of the first evidence of hMSCs differentiation is the formation of an extracellular matrix (ECM) where the collagen protein is its main component. This work demonstrated the multimodal CARS and SHG microscopy as a powerful non-invasive label free technique to investigate the collagen production dynamic in living cell 3D cultures. Its ability to image the cell morphology and the produced collagen distribution on a long term (4 weeks) experiment allowed to obtain important information about the cell-scaffold interaction and the ECM production. The very low limit reached in detecting collagen has permit...

  7. 3-D palinspastic restoration of normal faults in the Inner Moray Firth: implications for extensional basin development

    Science.gov (United States)

    Barr, David

    1985-10-01

    Balanced cross-section techniques, and the construction of a restored section, permit 2-dimensional palinspastic restorations to be made in both compressional and extensional terraines. In 3 dimensions, an equivalent restoration can be made by assuming conservation of bedding-plane area and considering the volume of a stratigraphic interval rather than its cross-sectional area. Extensional basins displaying upper crustal listric normal faulting are particularly amenable to this approach. Computerised 3-D restorations have been made of the Inner Moray Firth basin, offshore Scotland. This basin is not isostatically compensated, and was produced by 7-8% post-Triassic extension, of which 2.5-3% is post-Jurassic, above a detachment surface at 20-25 km depth, close to the base of the crust. Limited lower crustal thinning (and lithospheric stretching) has affected the eastern part of the basin, but this can account for no more than half of the measured upper crustal extension. Some of this shallow extension is probably coupled by low-angle faults or shear zones into major zones of lithospheric stretching such as the North Sea grabens, where it may help account for discrepancies between estimates of lithospheric thinning and upper crustal extension.

  8. Comparison of the transcriptomic profile of hepatic human induced pluripotent stem like cells cultured in plates and in a 3D microscale dynamic environment.

    Science.gov (United States)

    Leclerc, Eric; Kimura, Keiichi; Shinohara, Marie; Danoy, Mathieu; Le Gall, Morgane; Kido, Taketomo; Miyajima, Atsushi; Fujii, Teruo; Sakai, Yasuyuki

    2017-01-01

    We have compared the transcriptomic profiles of human induced pluripotent stem cells after their differentiation in hepatocytes like cells in plates and microfluidic biochips. The biochips provided a 3D and dynamic support during the cell differentiation when compared to the 2D static cultures in plates. The microarray have demonstrated the up regulation of important pathway related to liver development and maturation during the culture in biochips. Furthermore, the results of the transcriptomic profile, coupled with immunostaining, and RTqPCR analysis have shown typical biomarkers illustrating the presence of responders of biliary like cells, hepatocytes like cells, and endothelial like cells. However, the overall tissue still presented characteristic of immature and foetal patterns. Nevertheless, the biochip culture provided a specific micro-environment in which a complex multicellular differentiation toward liver could be oriented.

  9. Stress evolution during 3D single-layer visco-elastic buckle folding: Implications for the initiation of fractures

    Science.gov (United States)

    Liu, Xiaolong; Eckert, Andreas; Connolly, Peter

    2016-06-01

    Buckle folds of sedimentary strata commonly feature a variety of different fracture sets. Some fracture sets including outer arc tensile fractures and inner arc shear fractures at the fold hinge zones are well understood by the extensional and compressional strain/stress pattern. However, other commonly observed fracture sets, including tensile fractures parallel to the fold axis, tensile fractures cutting through the limb, extensional faults at the fold hinge, and other shear fractures of various orientations in the fold limb, fail to be intuitively explained by the strain/stress regimes during the buckling process. To obtain a better understanding of the conditions for the initiation of the various fractures sets associated with single-layer cylindrical buckle folds, a 3D finite element modeling approach using a Maxwell visco-elastic rheology is utilized. The influences of three model parameters with significant influence on fracture initiation are considered: burial depth, viscosity, and permeability. It is concluded that these parameters are critical for the initiation of major fracture sets at the hinge zone with varying degrees. The numerical simulation results further show that the buckling process fails to explain most of the fracture sets occurring in the limb unless the process of erosional unloading as a post-fold phenomenon is considered. For fracture sets that only develop under unrealistic boundary conditions, the results demonstrate that their development is realistic for a perclinal fold geometry. In summary, a more thorough understanding of fractures sets associated with buckle folds is obtained based on the simulation of in-situ stress conditions during the structural development of buckle folds.

  10. 3D MODELING OF GJ1214b's ATMOSPHERE: FORMATION OF INHOMOGENEOUS HIGH CLOUDS AND OBSERVATIONAL IMPLICATIONS

    Energy Technology Data Exchange (ETDEWEB)

    Charnay, B.; Meadows, V.; Misra, A.; Arney, G. [Astronomy Department, University of Washington, Seattle, WA 98125 (United States); Leconte, J., E-mail: bcharnay@uw.edu [Canadian Institute for Theoretical Astrophysics, 60 St George Street, University of Toronto, Toronto, ON M5S 3H8 (Canada)

    2015-11-01

    The warm sub-Neptune GJ1214b has a featureless transit spectrum that may be due to the presence of high and thick clouds or haze. Here, we simulate the atmosphere of GJ1214b with a 3D General Circulation Model for cloudy hydrogen-dominated atmospheres, including cloud radiative effects. We show that the atmospheric circulation is strong enough to transport micrometric cloud particles to the upper atmosphere and generally leads to a minimum of cloud at the equator. By scattering stellar light, clouds increase the planetary albedo to 0.4–0.6 and cool the atmosphere below 1 mbar. However, the heating by ZnS clouds leads to the formation of a stratospheric thermal inversion above 10 mbar, with temperatures potentially high enough on the dayside to evaporate KCl clouds. We show that flat transit spectra consistent with Hubble Space Telescope observations are possible if cloud particle radii are around 0.5 μm, and that such clouds should be optically thin at wavelengths >3 μm. Using simulated cloudy atmospheres that fit the observed spectra we generate transit, emission, and reflection spectra and phase curves for GJ1214b. We show that a stratospheric thermal inversion would be readily accessible in near- and mid-infrared atmospheric spectral windows. We find that the amplitude of the thermal phase curves is strongly dependent on metallicity, but only slightly impacted by clouds. Our results suggest that primary and secondary eclipses and phase curves observed by the James Webb Space Telescope in the near- to mid-infrared should provide strong constraints on the nature of GJ1214b's atmosphere and clouds.

  11. Establishment of a heterotypic 3D culture system to evaluate the interaction of TREG lymphocytes and NK cells with breast cancer.

    Science.gov (United States)

    Augustine, Tanya N; Dix-Peek, Thérèse; Duarte, Raquel; Candy, Geoffrey P

    2015-11-01

    Three-dimensional (3D) culture approaches to investigate breast tumour progression are yielding information more reminiscent of the in vivo microenvironment. We have established a 3D Matrigel system to determine the interactions of luminal phenotype MCF-7 cells and basal phenotype MDA-MB-231 cells with regulatory T lymphocytes and Natural Killer cells. Immune cells were isolated from peripheral blood using magnetic cell sorting and their phenotype validated using flow cytometry both before and after activation with IL-2 and phytohaemagglutinin. Following the establishment of the heterotypic culture system, tumour cells displayed morphologies and cell-cell associations distinct to that observed in 2D monolayer cultures, and associated with tissue remodelling and invasion processes. We found that the level of CCL4 secretion was influenced by breast cancer phenotype and immune stimulation. We further established that for RNA extraction, the use of proteinase K in conjunction with the Qiagen RNeasy Mini Kit and only off-column DNA digestion gave the best RNA yield, purity and integrity. We also investigated the efficacy of the culture system for immunolocalisation of the biomarkers oestrogen receptor-α and the glycoprotein mucin 1 in luminal phenotype breast cancer cells; and epidermal growth factor receptor in basal phenotype breast cancer cells, in formalin-fixed, paraffin-wax embedded cultures. The expression of these markers was shown to vary under immune mediation. We thus demonstrate the feasibility of using this co-culture system for downstream applications including cytokine analysis, immunolocalisation of tumour biomarkers on serial sections and RNA extraction in accordance with MIQE guidelines.

  12. Network dynamics of 3D engineered neuronal cultures: a new experimental model for in-vitro electrophysiology

    NARCIS (Netherlands)

    Frega, M.; Tedesco, M.; Massobrio, P.; Pesce, M.; Martinoia, S.

    2014-01-01

    Despite the extensive use of in-vitro models for neuroscientific investigations and notwithstanding the growing field of network electrophysiology, all studies on cultured cells devoted to elucidate neurophysiological mechanisms and computational properties, are based on 2D neuronal networks. These

  13. 3D seismic analysis of the Collyhurst Sandstone: implications for CO2 sequestration in the East Irish Sea Basin

    Science.gov (United States)

    Gamboa, Davide; Williams, John; Kirk, Karen; Gent, Christopher; Bentham, Michelle; Fellgett, Mark; Schofield, David

    2016-04-01

    Carbon Capture and Storage (CCS) is a vital technology towards low-carbon energy resources and the mitigation of global warming trends induced by rising CO2 levels in the atmosphere. The East Irish Sea Basin (EISB) is a key area for CCS in the western UK, having high CO2 storage potentials in explored hydrocarbon fields and in saline aquifers within the Permo-Triassic Sherwood Sandstone Formation. However, the theoretical storage potential of the EISB could be poorly estimated as the reservoir-prone Lower Permian formations are not considered in detail by current estimations. This work aims to fill this gap, focusing on the characterisation of the Lower Permian Collyhurst Sandstone Formation as a viable storage unit. The potential for CO2 storage is estimated as the total volume/area of suitable closures that are isolated by structural traps, occurring at depths suitable for CO2 injection and containment (>800m). Detailed structural and stratigraphic interpretations were made using 3D seismic data to assess the storage potential of the Collyhurst Sandstone Formation in the southern EISB. The basin strata is compartmentalised by numerous N-S trending faults. A higher degree of compartmentalisation occurs within regional anticlines where elongated tilted blocks are observed, bound by predominantly west-dipping faults that induce a variable offset of the Collyhurst Sandstone strata. Contrastingly, higher lateral continuity of this formation is observed within graben basins were faults are less frequent and with minor offset, thus potentially creating larger storage closures. Fault dip orientation in the grabens is variable, with west and east dipping faults occurring as a function of large east-dipping listric faults. This study was complemented by the stress modelling of the interpreted faults in order to assess the risk of CO2 leakage. Analysis of borehole breakouts observed in four approximately vertical wells in the EISB suggest a maximum horizontal stress

  14. Bioprinting of 3D hydrogels.

    Science.gov (United States)

    Stanton, M M; Samitier, J; Sánchez, S

    2015-08-07

    Three-dimensional (3D) bioprinting has recently emerged as an extension of 3D material printing, by using biocompatible or cellular components to build structures in an additive, layer-by-layer methodology for encapsulation and culture of cells. These 3D systems allow for cell culture in a suspension for formation of highly organized tissue or controlled spatial orientation of cell environments. The in vitro 3D cellular environments simulate the complexity of an in vivo environment and natural extracellular matrices (ECM). This paper will focus on bioprinting utilizing hydrogels as 3D scaffolds. Hydrogels are advantageous for cell culture as they are highly permeable to cell culture media, nutrients, and waste products generated during metabolic cell processes. They have the ability to be fabricated in customized shapes with various material properties with dimensions at the micron scale. 3D hydrogels are a reliable method for biocompatible 3D printing and have applications in tissue engineering, drug screening, and organ on a chip models.

  15. Mitigation of Lethal Radiation Syndrome in Mice by Intramuscular Injection of 3D Cultured Adherent Human Placental Stromal Cells.

    Directory of Open Access Journals (Sweden)

    Elena Gaberman

    Full Text Available Exposure to high lethal dose of ionizing radiation results in acute radiation syndrome with deleterious systemic effects to different organs. A primary target is the highly sensitive bone marrow and the hematopoietic system. In the current study C3H/HeN mice were total body irradiated by 7.7 Gy. Twenty four hrs and 5 days after irradiation 2×10(6 cells from different preparations of human derived 3D expanded adherent placental stromal cells (PLX were injected intramuscularly. Treatment with batches consisting of pure maternal cell preparations (PLX-Mat increased the survival of the irradiated mice from ∼27% to 68% (P<0.001, while cell preparations with a mixture of maternal and fetal derived cells (PLX-RAD increased the survival to ∼98% (P<0.0001. The dose modifying factor of this treatment for both 50% and 37% survival (DMF50 and DMF37 was∼1.23. Initiation of the more effective treatment with PLX-RAD injection could be delayed for up to 48 hrs after irradiation with similar effect. A delayed treatment by 72 hrs had lower, but still significantly effect (p<0.05. A faster recovery of the BM and improved reconstitution of all blood cell lineages in the PLX-RAD treated mice during the follow-up explains the increased survival of the cells treated irradiated mice. The number of CD45+/SCA1+ hematopoietic progenitor cells within the fast recovering population of nucleated BM cells in the irradiated mice was also elevated in the PLX-RAD treated mice. Our study suggests that IM treatment with PLX-RAD cells may serve as a highly effective "off the shelf" therapy to treat BM failure following total body exposure to high doses of radiation. The results suggest that similar treatments may be beneficial also for clinical conditions associated with severe BM aplasia and pancytopenia.

  16. Optimization of high grade glioma cell culture from surgical specimens for use in clinically relevant animal models and 3D immunochemistry.

    Science.gov (United States)

    Hasselbach, Laura A; Irtenkauf, Susan M; Lemke, Nancy W; Nelson, Kevin K; Berezovsky, Artem D; Carlton, Enoch T; Transou, Andrea D; Mikkelsen, Tom; deCarvalho, Ana C

    2014-01-07

    Glioblastomas, the most common and aggressive form of astrocytoma, are refractory to therapy, and molecularly heterogeneous. The ability to establish cell cultures that preserve the genomic profile of the parental tumors, for use in patient specific in vitro and in vivo models, has the potential to revolutionize the preclinical development of new treatments for glioblastoma tailored to the molecular characteristics of each tumor. Starting with fresh high grade astrocytoma tumors dissociated into single cells, we use the neurosphere assay as an enrichment method for cells presenting cancer stem cell phenotype, including expression of neural stem cell markers, long term self-renewal in vitro, and the ability to form orthotopic xenograft tumors. This method has been previously proposed, and is now in use by several investigators. Based on our experience of dissociating and culturing 125 glioblastoma specimens, we arrived at the detailed protocol we present here, suitable for routine neurosphere culturing of high grade astrocytomas and large scale expansion of tumorigenic cells for preclinical studies. We report on the efficiency of successful long term cultures using this protocol and suggest affordable alternatives for culturing dissociated glioblastoma cells that fail to grow as neurospheres. We also describe in detail a protocol for preserving the neurospheres 3D architecture for immunohistochemistry. Cell cultures enriched in CSCs, capable of generating orthotopic xenograft models that preserve the molecular signatures and heterogeneity of GBMs, are becoming increasingly popular for the study of the biology of GBMs and for the improved design of preclinical testing of potential therapies.

  17. A dynamic multi-organ-chip for long-term cultivation and substance testing proven by 3D human liver and skin tissue co-culture.

    Science.gov (United States)

    Wagner, Ilka; Materne, Eva-Maria; Brincker, Sven; Süssbier, Ute; Frädrich, Caroline; Busek, Mathias; Sonntag, Frank; Sakharov, Dmitry A; Trushkin, Evgeny V; Tonevitsky, Alexander G; Lauster, Roland; Marx, Uwe

    2013-09-21

    Current in vitro and animal tests for drug development are failing to emulate the systemic organ complexity of the human body and, therefore, to accurately predict drug toxicity. In this study, we present a multi-organ-chip capable of maintaining 3D tissues derived from cell lines, primary cells and biopsies of various human organs. We designed a multi-organ-chip with co-cultures of human artificial liver microtissues and skin biopsies, each a (1)/100,000 of the biomass of their original human organ counterparts, and have successfully proven its long-term performance. The system supports two different culture modes: i) tissue exposed to the fluid flow, or ii) tissue shielded from the underlying fluid flow by standard Transwell® cultures. Crosstalk between the two tissues was observed in 14-day co-cultures exposed to fluid flow. Applying the same culture mode, liver microtissues showed sensitivity at different molecular levels to the toxic substance troglitazone during a 6-day exposure. Finally, an astonishingly stable long-term performance of the Transwell®-based co-cultures could be observed over a 28-day period. This mode facilitates exposure of skin at the air-liquid interface. Thus, we provide here a potential new tool for systemic substance testing.

  18. Interactions between mesenchymal stem cells, adipocytes, and osteoblasts in a 3D tri-culture model of hyperglycemic conditions in the bone marrow microenvironment.

    Science.gov (United States)

    Rinker, Torri E; Hammoudi, Taymour M; Kemp, Melissa L; Lu, Hang; Temenoff, Johnna S

    2014-03-01

    Recent studies have found that uncontrolled diabetes and consequential hyperglycemic conditions can lead to an increased incidence of osteoporosis. Osteoblasts, adipocytes, and mesenchymal stem cells (MSCs) are all components of the bone marrow microenvironment and thus may have an effect on diabetes-related osteoporosis. However, few studies have investigated the influence of these three cell types on each other, especially in the context of hyperglycemia. Thus, we developed a hydrogel-based 3D culture platform engineered to allow live-cell retrieval in order to investigate the interactions between MSCs, osteoblasts, and adipocytes in mono-, co-, and tri-culture configurations under hyperglycemic conditions for 7 days of culture. Gene expression, histochemical analysis of differentiation markers, and cell viability were measured for all cell types, and MSC-laden hydrogels were degraded to retrieve cells to assess their colony-forming capacity. Multivariate models of gene expression data indicated that primary discrimination was dependent on the neighboring cell type, validating the need for co-culture configurations to study conditions modeling this disease state. MSC viability and clonogenicity were reduced when mono- and co-cultured with osteoblasts at high glucose levels. In contrast, MSCs showed no reduction of viability or clonogenicity when cultured with adipocytes under high glucose conditions, and the adipogenic gene expression indicates that cross-talk between MSCs and adipocytes may occur. Thus, our unique culture platform combined with post-culture multivariate analysis provided a novel insight into cellular interactions within the MSC microenvironment and highlights the necessity of multi-cellular culture systems for further investigation of complex pathologies such as diabetes and osteoporosis.

  19. Osteogenic potential of human adipose-tissue-derived mesenchymal stromal cells cultured on 3D-printed porous structured titanium.

    Science.gov (United States)

    Lewallen, Eric A; Jones, Dakota L; Dudakovic, Amel; Thaler, Roman; Paradise, Christopher R; Kremers, Hilal M; Abdel, Matthew P; Kakar, Sanjeev; Dietz, Allan B; Cohen, Robert C; Lewallen, David G; van Wijnen, Andre J

    2016-05-01

    Integration of porous metal prosthetics, which restore form and function of irreversibly damaged joints, into remaining healthy bone is critical for implant success. We investigated the biological properties of adipose-tissue-derived mesenchymal stromal/stem cells (AMSCs) and addressed their potential to alter the in vitro microenvironment of implants. We employed human AMSCs as a practical source for musculoskeletal applications because these cells can be obtained in large quantities, are multipotent, and have trophic paracrine functions. AMSCs were cultured on surgical-grade porous titanium disks as a model for orthopedic implants. We monitored cell/substrate attachment, cell proliferation, multipotency, and differentiation phenotypes of AMSCs upon osteogenic induction. High-resolution scanning electron microscopy and histology revealed that AMSCs adhere to the porous metallic surface. Compared to standard tissue culture plastic, AMSCs grown in the porous titanium microenvironment showed differences in temporal expression for genes involved in cell cycle progression (CCNB2, HIST2H4), extracellular matrix production (COL1A1, COL3A1), mesenchymal lineage identity (ACTA2, CD248, CD44), osteoblastic transcription factors (DLX3, DLX5, ID3), and epigenetic regulators (EZH1, EZH2). We conclude that metal orthopedic implants can be effectively seeded with clinical-grade stem/stromal cells to create a pre-conditioned implant.

  20. Survival and growth of isolated pre-antral follicles from human ovarian medulla tissue during long-term 3D culture

    DEFF Research Database (Denmark)

    Yin, H L; Kristensen, S G; Jiang, H

    2016-01-01

    system for more than 30 days. WHAT IS KNOWN ALREADY: Ovarian tissue cryopreservation followed by transplantation is a promising fertility preservation approach for cancer patients. However, transplantation of cryopreserved tissue to patients may carry the risk of re-implanting malignant cells. Grafting...... that is normally discarded, possess a developmental potential which may be used to devise safer fertility preservation methods for patients who are at high risk of malignant contamination of their ovarian tissue. STUDY FUNDING/COMPETING INTERESTS: The Child Cancer Foundation in Denmark (2012-26) and the EU......STUDY QUESTION: Can human pre-antral follicles isolated enzymatically from surplus medulla tissue survive and grow in vitro during long-term 3D culture? SUMMARY ANSWER: Secondary human follicles can develop to small antral follicles and remain hormonally active in an alginate-encapsulation culture...

  1. Culture and establishment of self-renewing human and mouse adult liver and pancreas 3D organoids and their genetic manipulation.

    Science.gov (United States)

    Broutier, Laura; Andersson-Rolf, Amanda; Hindley, Christopher J; Boj, Sylvia F; Clevers, Hans; Koo, Bon-Kyoung; Huch, Meritxell

    2016-09-01

    Adult somatic tissues have proven difficult to expand in vitro, largely because of the complexity of recreating appropriate environmental signals in culture. We have overcome this problem recently and developed culture conditions for adult stem cells that allow the long-term expansion of adult primary tissues from small intestine, stomach, liver and pancreas into self-assembling 3D structures that we have termed 'organoids'. We provide a detailed protocol that describes how to grow adult mouse and human liver and pancreas organoids, from cell isolation and long-term expansion to genetic manipulation in vitro. Liver and pancreas cells grow in a gel-based extracellular matrix (ECM) and a defined medium. The cells can self-organize into organoids that self-renew in vitro while retaining their tissue-of-origin commitment, genetic stability and potential to differentiate into functional cells in vitro (hepatocytes) and in vivo (hepatocytes and endocrine cells). Genetic modification of these organoids opens up avenues for the manipulation of adult stem cells in vitro, which could facilitate the study of human biology and allow gene correction for regenerative medicine purposes. The complete protocol takes 1-4 weeks to generate self-renewing 3D organoids and to perform genetic manipulation experiments. Personnel with basic scientific training can conduct this protocol.

  2. Feasibility of using sodium chloride as a tracer for the characterization of the distribution of matter in complex multi-compartment 3D bioreactors for stem cell culture.

    Science.gov (United States)

    Gerlach, Jörg C; Witaschek, Tom; Strobel, Catrin; Brayfield, Candace A; Bornemann, Reinhard; Catapano, Gerardo; Zeilinger, Katrin

    2010-06-01

    The experimental characterization of the distribution of matter in complex multi-compartment three-dimensional membrane bioreactors for human cell culture is complicated by tracer interactions with the membranes and other bioreactor constituents. This is due to the fact that membranes with a high specific surface area often feature a hydrophobic chemical backbone that may adsorb tracers often used to this purpose, such as proteins and dyes. Membrane selectivity, and its worsening caused by protein adsorption, may also hinder tracer transfer across neighboring compartments, thus preventing effective characterization of the distribution of matter in the whole bioreactor. Tracer experiments with sodium chloride (NaCl) may overcome some of these limitations and be effectively used to characterize the distribution of matter in complex 3D multi-compartments membrane bioreactors for stem cell culture. NaCl freely permeates most used membranes, it does not adsorb on uncharged membranes, and its concentration may be accurately measured in terms of solution conductivity. In this preliminary study, the feasibility of complex multi-compartment membrane bioreactors was investigated with a NaCl concentration pulse challenge to characterize how their distribution of matter changes when they are operated under different conditions. In particular, bioreactors consisting of three different membrane types stacked on top of one another to form a 3D network were characterized under different feed conditions.

  3. 3D video

    CERN Document Server

    Lucas, Laurent; Loscos, Céline

    2013-01-01

    While 3D vision has existed for many years, the use of 3D cameras and video-based modeling by the film industry has induced an explosion of interest for 3D acquisition technology, 3D content and 3D displays. As such, 3D video has become one of the new technology trends of this century.The chapters in this book cover a large spectrum of areas connected to 3D video, which are presented both theoretically and technologically, while taking into account both physiological and perceptual aspects. Stepping away from traditional 3D vision, the authors, all currently involved in these areas, provide th

  4. 3D Animation Essentials

    CERN Document Server

    Beane, Andy

    2012-01-01

    The essential fundamentals of 3D animation for aspiring 3D artists 3D is everywhere--video games, movie and television special effects, mobile devices, etc. Many aspiring artists and animators have grown up with 3D and computers, and naturally gravitate to this field as their area of interest. Bringing a blend of studio and classroom experience to offer you thorough coverage of the 3D animation industry, this must-have book shows you what it takes to create compelling and realistic 3D imagery. Serves as the first step to understanding the language of 3D and computer graphics (CG)Covers 3D anim

  5. 3D-CSIA: carbon, chlorine, and hydrogen isotope fractionation in transformation of TCE to ethene by a Dehalococcoides culture.

    Science.gov (United States)

    Kuder, Tomasz; van Breukelen, Boris M; Vanderford, Mindy; Philp, Paul

    2013-09-03

    Carbon (C), chlorine (Cl), and hydrogen (H) isotope effects were determined during dechlorination of TCE to ethene by a mixed Dehalococcoides (Dhc) culture. The C isotope effects for the dechlorination steps were consistent with data published in the past for reductive dechlorination (RD) by Dhc. The Cl effects (combined with an inverse H effect in TCE) suggested that dechlorination proceeded through nucleophilic reactions with cobalamin rather than by an electron transfer mechanism. Depletions of (37)Cl in daughter compounds, resulting from fractionation at positions away from the dechlorination center (secondary isotope effects), further support the nucleophilic dechlorination mechanism. Determination of C and Cl isotope ratios of the reactants and products in the reductive dechlorination chain offers a potential tool for differentiation of Dhc activity from alternative transformation mechanisms (e.g., aerobic degradation and reductive dechlorination proceeding via outer sphere mechanisms), in studies of in situ attenuation of chlorinated ethenes. Hydrogenation of the reaction products (DCE, VC, and ethene) showed a major preference for the (1)H isotope. Detection of depleted dechlorination products could provide a line of evidence in discrimination between alternative sources of TCE (e.g., evolution from DNAPL sources or from conversion of PCE).

  6. Novel Image Analysis Approach Quantifies Morphological Characteristics of 3D Breast Culture Acini with Varying Metastatic Potentials

    Directory of Open Access Journals (Sweden)

    Lindsey McKeen Polizzotti

    2012-01-01

    Full Text Available Prognosis of breast cancer is primarily predicted by the histological grading of the tumor, where pathologists manually evaluate microscopic characteristics of the tissue. This labor intensive process suffers from intra- and inter-observer variations; thus, computer-aided systems that accomplish this assessment automatically are in high demand. We address this by developing an image analysis framework for the automated grading of breast cancer in in vitro three-dimensional breast epithelial acini through the characterization of acinar structure morphology. A set of statistically significant features for the characterization of acini morphology are exploited for the automated grading of six (MCF10 series cell line cultures mimicking three grades of breast cancer along the metastatic cascade. In addition to capturing both expected and visually differentiable changes, we quantify subtle differences that pose a challenge to assess through microscopic inspection. Our method achieves 89.0% accuracy in grading the acinar structures as nonmalignant, noninvasive carcinoma, and invasive carcinoma grades. We further demonstrate that the proposed methodology can be successfully applied for the grading of in vivo tissue samples albeit with additional constraints. These results indicate that the proposed features can be used to describe the relationship between the acini morphology and cellular function along the metastatic cascade.

  7. In Vivo 3D Analysis of Thoracic Kinematics: Changes in Size and Shape During Breathing and Their Implications for Respiratory Function in Recent Humans and Fossil Hominins.

    Science.gov (United States)

    Bastir, Markus; García-Martínez, Daniel; Torres-Tamayo, Nicole; Sanchis-Gimeno, Juan Alberto; O'Higgins, Paul; Utrilla, Cristina; Torres Sánchez, Isabel; García Río, Francisco

    2017-02-01

    The human ribcage expands and contracts during respiration as a result of the interaction between the morphology of the ribs, the costo-vertebral articulations and respiratory muscles. Variations in these factors are said to produce differences in the kinematics of the upper thorax and the lower thorax, but the extent and nature of any such differences and their functional implications have not yet been quantified. Applying geometric morphometrics we measured 402 three-dimensional (3D) landmarks and semilandmarks of 3D models built from computed tomographic scans of thoraces of 20 healthy adult subjects in maximal forced inspiration (FI) and expiration (FE). We addressed the hypothesis that upper and lower parts of the ribcage differ in kinematics and compared different models of functional compartmentalization. During inspiration the thorax superior to the level of the sixth ribs undergoes antero-posterior expansion that differs significantly from the medio-lateral expansion characteristic of the thorax below this level. This supports previous suggestions for dividing the thorax into a pulmonary and diaphragmatic part. While both compartments differed significantly in mean size and shape during FE and FI the size changes in the lower compartment were significantly larger. Additionally, for the same degree of kinematic shape change, the pulmonary thorax changes less in size than the diaphragmatic thorax. Therefore, variations in the form and function of the diaphragmatic thorax will have a strong impact on respiratory function. This has important implications for interpreting differences in thorax shape in terms of respiratory functional differences within and among recent humans and fossil hominins. Anat Rec, 300:255-264, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  8. AlgiMatrix™-Based 3D Cell Culture System as an In Vitro Tumor Model: An Important Tool in Cancer Research.

    Science.gov (United States)

    Godugu, Chandraiah; Singh, Mandip

    2016-01-01

    Routinely used two-dimensional cell culture-based models often fail while translating the observations into in vivo models. This setback is more common in cancer research, due to several reasons. The extracellular matrix and cell-to-cell interactions are not present in two-dimensional (2D) cell culture models. Diffusion of drug molecules into cancer cells is hindered by barriers of extracellular components in in vivo conditions, these barriers are absent in 2D cell culture models. To better mimic or simulate the in vivo conditions present in tumors, the current study used the alginate based three-dimensional cell culture (AlgiMatrix™) model, which resembles close to the in vivo tumor models. The current study explains the detailed protocols involved in AlgiMatrix™ based in vitro non-small-cell lung cancer (NSCLC) models. The suitability of this model was studied by evaluating, cytotoxicity, apoptosis, and penetration of nanoparticles into the in vitro tumor spheroids. This study also demonstrated the effect of EphA2 receptor targeted docetaxel-loaded nanoparticles on MDA-MB-468 TNBC cell lines. The methods section is subdivided into three subsections such as (1) preparation of AlgiMatrix™-based 3D in vitro tumor models and cytotoxicity assays, (2) free drug and nanoparticle uptake into spheroid studies, and (3) western blot, IHC, and RT-PCR studies.

  9. PPO/PEO modified hollow fiber membranes improved sensitivity of 3D cultured hepatocytes to drug toxicity via suppressing drug adsorption on membranes.

    Science.gov (United States)

    Shen, Chong; Meng, Qin; He, Wenjuan; Wang, Qichen; Zhang, Guoliang

    2014-11-01

    The three dimensional (3D) cell culture in polymer-based micro system has become a useful tool for in vitro drug discovery. Among those polymers, polysulfone hollow fiber membrane (PSf HFM) is commonly used to create a microenvironment for cells. However, the target drug may adsorb on the polymeric surface, and this elicits negative impacts on cell exposure due to the reduced effective drug concentration in culture medium. In order to reduce the drug adsorption, PSf membrane were modified with hydrophilic Pluronic (PEO-b-PPO-b-PEO) copolymers, L121, P123 and F127 (PEO contents increase from 10%, 30% to 70%), by physical adsorption. As a result, the hydrophilicity of HFMs increased at an order of PSfF127>P123>L121 HFMs. The three modified membrane all showed significant resistance to adsorption of acid/neutral drugs. More importantly, the adsorption of base drugs were largely reduced to an average value of 11% on the L121 HFM. The improved resistance to drug adsorption could be attributed to the synergy of hydrophobic/neutrally charged PPO and hydrophilic PEO. The L121 HFM was further assessed by evaluating the drug hepatotoxicity in 3D culture of hepatocytes. The base drugs, clozapine and doxorubicin, showed more sensitive hepatotoxicity on hepatocytes in L121 HFM than in PSf HFM, while the acid drug, salicylic acid, showed the similar hepatotoxicity to hepatocytes in both HFMs. Our finding suggests that PSf HFM modified by PEO-b-PPO-b-PEO copolymers can efficiently resist the drug adsorption onto polymer membrane, and consequently improve the accuracy and sensitivity of in vitro hepatotoxic drug screening.

  10. The role of hair follicle nestin-expressing stem cells during whisker sensory-nerve growth in long-term 3D culture.

    Science.gov (United States)

    Mii, Sumiyuki; Duong, Jennifer; Tome, Yasunori; Uchugonova, Aisada; Liu, Fang; Amoh, Yasuyuki; Saito, Norimitsu; Katsuoka, Kensei; Hoffman, Robert M

    2013-07-01

    We have previously reported that nestin-expressing hair follicle stem cells can differentiate into neurons, Schwann cells, and other cell types. In the present study, vibrissa hair follicles, including their sensory nerve stump, were excised from transgenic mice in which the nestin promoter drives green fluorescent protein (ND-GFP mice), and were placed in 3D histoculture supported by Gelfoam®. β-III tubulin-positive fibers, consisting of ND-GFP-expressing cells, extended up to 500 µm from the whisker nerve stump in histoculture. The growing fibers had growth cones on their tips expressing F-actin. These findings indicate that β-III tubulin-positive fibers elongating from the whisker follicle sensory nerve stump were growing axons. The growing whisker sensory nerve was highly enriched in ND-GFP cells which appeared to play a major role in its elongation and interaction with other nerves in 3D culture, including the sciatic nerve, the trigeminal nerve, and the trigeminal nerve ganglion. The results of the present report suggest a major function of the nestin-expressing stem cells in the hair follicle is for growth of the follicle sensory nerve. Copyright © 2013 Wiley Periodicals, Inc.

  11. On Dynamic Characteristics of Culture and its Implications

    Institute of Scientific and Technical Information of China (English)

    赵凌志

    2015-01-01

    This paper mainly discusses the dynamic characteristic of western culture and Chinese culture from the intercultural perspective.Then it puts forward some implications for English teaching,it indicates that English teachers should pay due attention to improve the students cultural awareness and their own cultural teaching ability,adjust teaching content to adapt them to cultural changes.

  12. 基于Kinect体感交互技术的3D服饰文化展示系统%Kinect-based 3D Costume Culture Display System

    Institute of Scientific and Technical Information of China (English)

    姜延; 马文轩; 陈剑华; 刘正东

    2015-01-01

    To carry forward traditional costume culture, a Kinect-based 3D costume culture display system was designed based on somatosensory interactive technology and Augmented Reality (AR( technology. Different from virtual mirror for commercial use, the system pays more attention to the popularization of Chinese traditional costume culture. The costumes, which are displayed under several themes such as Ancient Costumes, Ethnic Costumes and Costumes for the Olympic Games, would give viewers a completely new experience.%为弘扬传统服饰文化,本文融合体感交互技术与增强现实技术,设计并实现了基于Kinect体感设备的3D服饰文化展示系统。不同于以往商业行为中的虚拟试衣镜,系统更注重中华传统服饰文化的普及,包括国粹箐华历代服饰、少数民族服饰、奥运主题服饰在内的多个主题将给使用者带来全新的用户体验。

  13. Responsiveness to PI3K and MEK inhibitors in breast cancer. Use of a 3D culture system to study pathways related to hormone independence in mice.

    Directory of Open Access Journals (Sweden)

    Maria Laura Polo

    Full Text Available BACKGROUND: A significant proportion of breast cancer patients face failure of endocrine therapy due to the acquisition of endocrine resistance. We have explored mechanisms involved in such disease progression by using a mouse breast cancer model that is induced by medroxyprogesterone acetate (MPA. These tumors transit through different stages of hormone sensitivity. However, when cells from tumor variants were seeded on plastic, all were stimulated by progestins and inhibited by antiprogestins such as RU486. Furthermore, cells from a RU486-resistant tumor variant recovered antiprogestin sensitivity. HYPOTHESIS: A three-dimensional (3D culture system, by maintaining differential cellular organization that is typical of each tumor variant, may allow for the maintenance of particular hormone responses and thus be appropriate for the study of the effects of specific inhibitors of signaling pathways associated with disease progression. METHOD: We compared the behavior of tumors growing in vivo and cancer cells ex vivo (in 3D Matrigel. In this system, we evaluated the effects of kinase inhibitors and hormone antagonists on tumor growth. PRINCIPAL FINDINGS: LY294002, a PI3K/AKT pathway inhibitor, decreased both tumor growth in vivo and cell survival in Matrigel in MPA-independent tumors with higher AKT activity. Induction of cell death by anti-hormones such as ICI182780 and ZK230211 was more effective in MPA-dependent tumors with lower AKT activity. Inhibition of MEK with PD98059 did not affect tumor growth in any tested variant. Finally, while Matrigel reproduced differential responsiveness of MPA-dependent and -independent breast cancer cells, it was not sufficient to preserve antiprogestin resistance of RU486-resistant tumors. CONCLUSION: We demonstrated that the PI3K/AKT pathway is relevant for MPA-independent tumor growth. Three-dimensional cultures were useful to test the effects of kinase inhibitors on breast cancer growth and highlight the

  14. A new back-and-forth iterative method for time-reversed convection modeling: Implications for the Cenozoic evolution of 3-D structure and dynamics of the mantle

    Science.gov (United States)

    Glišović, Petar; Forte, Alessandro M.

    2016-06-01

    The 3-D distribution of buoyancy in the convecting mantle drives a suite of convection-related manifestations. Although seismic tomography is providing increasingly resolved images of the present-day mantle heterogeneity, the distribution of mantle density variations in the geological past is unknown, and, by implication, this is true for the convection-related observables. The one major exception is tectonic plate motions, since geologic data are available to estimate their history and they currently provide the only available constraints on the evolution of 3-D mantle buoyancy in the past. We developed a new back-and-forth iterative method for time-reversed convection modeling with a procedure for matching plate velocity data at different instants in the past. The crucial aspect of this reconstruction methodology is to ensure that at all times plates are driven by buoyancy forces in the mantle and not vice versa. Employing tomography-based retrodictions over the Cenozoic, we estimate the global amplitude of the following observables: dynamic surface topography, the core-mantle boundary ellipticity, the free-air gravity anomalies, and the global divergence rates of tectonic plates. One of the major benefits of the new data assimilation method is the stable recovery of much shorter wavelength changes in heterogeneity than was possible in our previous work. We now resolve what appears to be two-stage subduction of the Farallon plate under the western U.S. and a deeply rooted East African Plume that is active under the Ethiopian volcanic fields during the Early Eocene.

  15. EUROPEANA AND 3D

    Directory of Open Access Journals (Sweden)

    D. Pletinckx

    2012-09-01

    Full Text Available The current 3D hype creates a lot of interest in 3D. People go to 3D movies, but are we ready to use 3D in our homes, in our offices, in our communication? Are we ready to deliver real 3D to a general public and use interactive 3D in a meaningful way to enjoy, learn, communicate? The CARARE project is realising this for the moment in the domain of monuments and archaeology, so that real 3D of archaeological sites and European monuments will be available to the general public by 2012. There are several aspects to this endeavour. First of all is the technical aspect of flawlessly delivering 3D content over all platforms and operating systems, without installing software. We have currently a working solution in PDF, but HTML5 will probably be the future. Secondly, there is still little knowledge on how to create 3D learning objects, 3D tourist information or 3D scholarly communication. We are still in a prototype phase when it comes to integrate 3D objects in physical or virtual museums. Nevertheless, Europeana has a tremendous potential as a multi-facetted virtual museum. Finally, 3D has a large potential to act as a hub of information, linking to related 2D imagery, texts, video, sound. We describe how to create such rich, explorable 3D objects that can be used intuitively by the generic Europeana user and what metadata is needed to support the semantic linking.

  16. Clinical implication of parameteroptimized 3D-FISP MR angiography (MRA) in children with aortic coarctation: comparison with catheter angiography; Stellenwert einer parameteroptimierten 3D-FISP-MR-Angiographie bei Kindern mit Aortenisthmusstenose im Vergleich zur Katheterangiographie

    Energy Technology Data Exchange (ETDEWEB)

    Kramer, U.; Dammann, F.; Miller, S.; Fenchel, M.; Claussen, C.D. [Universitaet Tuebingen (Germany). Abteilung Radiologische Diagnostik; Greil, G.; Sieverding, L. [Universitaet Tuebingen (Germany). Abteilung Kinderkardiologie; Schick, F. [Universitaet Tuebingen (Germany). Sektion Experimentelle Radiologie

    2004-10-01

    Purpose: To implement parameter-optimized 3D-FISP MR angiography (MRA) with interleaved double-slab excitation and to compare the result with catheter angiography in children with aortic coarctation. Materials and Methods: Eighteen children aged 2-15 years (mean 9.1 years) underwent MR imaging on a 1.5 T body scanner (Magnetom Vision, Siemens, Germany). All patients had undergone correlative catheter angiography. T1-weighted turbo spin echo (TSE) images (TR 600 ms, TE 17 ms, flip 160 , slice thickness 2-4 mm) were obtained in axial and parasagittal orientation, followed by an optimized 3D-FISP MR angiography in a sagittal plane (TR 12.5 ms, TE 5.5 ms, flip 22 , matrix 256 x 256, slice thickness 1.25 mm). All children were sedated but on spontaneous breathing. Image quality was graded by two experienced reviewers using a 4-point scoring system. Source images and reformatted maximum intensity projections (MIP) were analyzed for blood-tissue contrast as well as size and focal stenoses of the aortic arch. Results: Aortic coarctation was found in 13 of 18 patients, using the 3D-FISP MRA. A high correlation value (r=0.96) was found compared to catheter angiography. Image quality was high in 94% with well defined blood-tissue contrast in all cases. The sensitivity of flow and breathing motion was low. Examination time was about 15 minutes depending on volume of interest and heart rate. Diagnostic accuracy has shown improvement using a combined analysis of source and MIP images. The mentioned technique has provided an excellent display of thoracic vasculature. (orig.)

  17. 3D GEOMATICS TECHNIQUES FOR AN INTEGRATED APPROACH TO CULTURAL HERITAGE KNOWLEDGE: THE CASE OF SAN MICHELE IN ACERBOLI’S CHURCH IN SANTARCANGELO DI ROMAGNA

    Directory of Open Access Journals (Sweden)

    G. Bitelli

    2017-05-01

    Full Text Available The modern Geomatics techniques, such as Terrestrial Laser Scanner (TLS and multi-view Structure from Motion (SfM, are gaining more and more interest in the Cultural Heritage field. All the data acquired with these technologies could be stored and managed together with other information in a Historical Building Information Model (HBIM. In this paper, it will be shown the case study of the San Michele in Acerboli’s church, located in Santarcangelo di Romagna, Italy. This church, dated about the 6th century A.D., represents a high relevant Romanic building of the high Medieval period. The building presents an irregular square plan with a different length of the lateral brick walls and a consequential oblique one in correspondence of the apse. Nevertheless, the different lengths of the lateral brick walls are balanced thanks to the irregular spaces between the windows. Different changes occurred during the centuries, such as the closing of the seven main doors and the building of the bell tower, in the 11th century A.D., which is nowadays the main entrance of the church. An integrated survey was realized, covering the exterior and the interior. The final 3D model represents a valid support not only for documentation, but also to maintain and manage in an integrate approach the available knowledge of this Cultural Heritage site, developing a HBIM system in which all the mentioned historical, geometrical, material matters are collected.

  18. IZDELAVA TISKALNIKA 3D

    OpenAIRE

    Brdnik, Lovro

    2015-01-01

    Diplomsko delo analizira trenutno stanje 3D tiskalnikov na trgu. Prikazan je razvoj in principi delovanja 3D tiskalnikov. Predstavljeni so tipi 3D tiskalnikov, njihove prednosti in slabosti. Podrobneje je predstavljena zgradba in delovanje koračnih motorjev. Opravljene so meritve koračnih motorjev. Opisana je programska oprema za rokovanje s 3D tiskalniki in komponente, ki jih potrebujemo za izdelavo. Diploma se oklepa vprašanja, ali je izdelava 3D tiskalnika bolj ekonomična kot pa naložba v ...

  19. 3D and Education

    Science.gov (United States)

    Meulien Ohlmann, Odile

    2013-02-01

    Today the industry offers a chain of 3D products. Learning to "read" and to "create in 3D" becomes an issue of education of primary importance. 25 years professional experience in France, the United States and Germany, Odile Meulien set up a personal method of initiation to 3D creation that entails the spatial/temporal experience of the holographic visual. She will present some different tools and techniques used for this learning, their advantages and disadvantages, programs and issues of educational policies, constraints and expectations related to the development of new techniques for 3D imaging. Although the creation of display holograms is very much reduced compared to the creation of the 90ies, the holographic concept is spreading in all scientific, social, and artistic activities of our present time. She will also raise many questions: What means 3D? Is it communication? Is it perception? How the seeing and none seeing is interferes? What else has to be taken in consideration to communicate in 3D? How to handle the non visible relations of moving objects with subjects? Does this transform our model of exchange with others? What kind of interaction this has with our everyday life? Then come more practical questions: How to learn creating 3D visualization, to learn 3D grammar, 3D language, 3D thinking? What for? At what level? In which matter? for whom?

  20. Inhibition of MMP-2-mediated cellular invasion by NF-κB inhibitor DHMEQ in 3D culture of breast carcinoma MDA-MB-231 cells: A model for early phase of metastasis.

    Science.gov (United States)

    Ukaji, Tamami; Lin, Yinzhi; Okada, Shoshiro; Umezawa, Kazuo

    2017-02-08

    The three-dimensional (3D) culture of cancer cells provides an environmental condition closely related to the condition in vivo. It would especially be an ideal model for the early phase of metastasis, including the detachment and invasion of cancer cells from the primary tumor. In one hand, dehydroxymethylepoxyquinomicin (DHMEQ), an NF-κB inhibitor, is known to inhibit cancer progression and late phase metastasis in animal experiments. In the present research, we studied the inhibitory activity on the 3D invasion of breast carcinoma cells. Breast carcinoma MDA-MB-231 cells showed the most active invasion from spheroid among the cell lines tested. DHMEQ inhibited the 3D invasion of cells at the 3D-nontoxic concentrations. The PCR array analysis using RNA isolated from the 3D on-top cultured cells indicated that matrix metalloproteinase (MMP)-2 expression is lowered by DHMEQ. Knockdown of MMP-2 and an MMP inhibitor, GM6001, both inhibited the invasion. DHMEQ was shown to inhibit the promoter activity of MMP-2 in the reporter assay. Thus, DHMEQ was shown to inhibit NF-κB/MMP-2-dependent cellular invasion in 3D-cultured MDA-MB-231 cells, suggesting that DHMEQ would inhibit the early phase of metastasis.

  1. Expression of transcription factors after short-term exposure of Arabidopsis thaliana cell cultures to hypergravity and simulated microgravity (2-D/3-D clinorotation, magnetic levitation)

    Science.gov (United States)

    Babbick, M.; Dijkstra, C.; Larkin, O. J.; Anthony, P.; Davey, M. R.; Power, J. B.; Lowe, K. C.; Cogoli-Greuter, M.; Hampp, R.

    Gravity is an important environmental factor that controls plant growth and development. Studies have shown that the perception of gravity is not only a property of specialized cells, but can also be performed by undifferentiated cultured cells. In this investigation, callus of Arabidopsis thaliana cv. Columbia was used to investigate the initial steps of gravity-related signalling cascades, through altered expression of transcription factors (TFs). TFs are families of small proteins that regulate gene expression by binding to specific promoter sequences. Based on microarray studies, members of the gene families WRKY, MADS-box, MYB, and AP2/EREBP were selected for investigation, as well as members of signalling chains, namely IAA 19 and phosphoinositol-4-kinase. Using qRT-PCR, transcripts were quantified within a period of 30 min in response to hypergravity (8 g), clinorotation [2-D clinostat and 3-D random positioning machine (RPM)] and magnetic levitation (ML). The data indicated that (1) changes in gravity induced stress-related signalling, and (2) exposure in the RPM induced changes in gene expression which resemble those of magnetic levitation. Two dimensional clinorotation resulted in responses similar to those caused by hypergravity. It is suggested that RPM and ML are preferable to simulate microgravity than clinorotation.

  2. 3D culture of tonsil-derived mesenchymal stem cells in poly(ethylene glycol)-poly(L-alanine-co-L-phenyl alanine) thermogel.

    Science.gov (United States)

    Park, Min Hee; Yu, Yeonsil; Moon, Hyo Jung; Ko, Du Young; Kim, Han Su; Lee, Hyukjin; Ryu, Kyung Ha; Jeong, Byeongmoon

    2014-11-01

    Poly(ethylene glycol)-poly(L-alanine-co-L-phenyl alanine) (PEG-PAF) aqueous solutions undergo sol-to-gel transition as the temperature increases. The transition is driven by the micelle aggregation involving the partial dehydration of the PEG block and the partial increase in β-sheet content of the PAF block. Tonsil-tissue-derived mesenchymal stem cells (TMSCs), a new stem cell resource, are encapsulated through the sol-to-gel transition of the TMSC-suspended PEG-PAF aqueous solutions. The encapsulated TMSCs are in vitro 3D cultured by using induction media supplemented with adipogenic, osteogenic, or chondrogenic factors, where the TMSCs preferentially undergo chondrogenesis with high expressions of type II collagen and sulfated glycosaminoglycan. As a feasibility study of the PEG-PAF thermogel for injectable tissue engineering, the TMSCs encapsulated in hydrogels are implanted in the subcutaneous layer of mice by injecting the TMSC suspended PEG-PAF aqueous solution. The in vivo studies also prove that TMSCs undergo chondrogenesis with high expression of the chondrogenic biomarkers. This study suggests that the TMSCs can be an excellent resource of MSCs, and the thermogelling PEG-PAF is a promising injectable tissue engineering scaffold, particularly for chondrogenic differentiation of the stem cells. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Dextran and Polymer Polyethylene Glycol (PEG Coating Reduce Both 5 and 30 nm Iron Oxide Nanoparticle Cytotoxicity in 2D and 3D Cell Culture

    Directory of Open Access Journals (Sweden)

    Alisa Morss Clyne

    2012-05-01

    Full Text Available Superparamagnetic iron oxide nanoparticles are widely used in biomedical applications, yet questions remain regarding the effect of nanoparticle size and coating on nanoparticle cytotoxicity. In this study, porcine aortic endothelial cells were exposed to 5 and 30 nm diameter iron oxide nanoparticles coated with either the polysaccharide, dextran, or the polymer polyethylene glycol (PEG. Nanoparticle uptake, cytotoxicity, reactive oxygen species (ROS formation, and cell morphology changes were measured. Endothelial cells took up nanoparticles of all sizes and coatings in a dose dependent manner, and intracellular nanoparticles remained clustered in cytoplasmic vacuoles. Bare nanoparticles in both sizes induced a more than 6 fold increase in cell death at the highest concentration (0.5 mg/mL and led to significant cell elongation, whereas cell viability and morphology remained constant with coated nanoparticles. While bare 30 nm nanoparticles induced significant ROS formation, neither 5 nm nanoparticles (bare or coated nor 30 nm coated nanoparticles changed ROS levels. Furthermore, nanoparticles were more toxic at lower concentrations when cells were cultured within 3D gels. These results indicate that both dextran and PEG coatings reduce nanoparticle cytotoxicity, however different mechanisms may be important for different size nanoparticles.

  4. Comparison of osteoclastogenesis and resorption activity of human osteoclasts on tissue culture polystyrene and on natural extracellular bone matrix in 2D and 3D.

    Science.gov (United States)

    Kleinhans, C; Schmid, F F; Schmid, F V; Kluger, P J

    2015-07-10

    Bone homeostasis is maintained by osteoblasts (bone formation) and osteoclasts (bone resorption). While there have been numerous studies investigating mesenchymal stem cells and their potential to differentiate into osteoblasts as well as their interaction with different bone substitute materials, there is only limited knowledge concerning in vitro generated osteoclasts. Due to the increasing development of degradable bone-grafting materials and the need of sophisticated in vitro test methods, it is essential to gain deeper insight into the process of osteoclastogenesis and the resorption functionality of human osteoclasts. Therefore, we focused on the comparison of osteoclastogenesis and resorption activity on tissue culture polystyrene (TCPS) and bovine extracellular bone matrices (BMs). Cortical bone slices were used as two-dimensional (2D) substrates, whereas a thermally treated cancellous bone matrix was used for three-dimensional (3D) experiments. We isolated primary human monocytes and induced osteoclastogenesis by medium supplementation. Subsequently, the expression of the vitronectin receptor (αVβ3) and cathepsin K as well as the characteristic actin formation on TCPS and the two BMs were examined. The cell area of human osteoclasts was analyzed on TCPS and on BMs, whereas significantly larger osteoclasts could be detected on BMs. Additionally, we compared the diameter of the sealing zones with the measured diameter of the resorption pits on the BMs and revealed similar diameters of the sealing zones and the resorption pits. We conclude that using TCPS as culture substrate does not affect the expression of osteoclast-specific markers. The analysis of resorption activity can successfully be conducted on cortical as well as on cancellous bone matrices. For new in vitro test systems concerning bone resorption, we suggest the establishment of a 2D assay for high throughput screening of new degradable bone substitute materials with osteoclasts.

  5. 3D digitization of mosaics

    Directory of Open Access Journals (Sweden)

    Anna Maria Manferdini

    2012-11-01

    Full Text Available In this paper we present a methodology developed to access to Cultural Heritage information using digital 3d reality-based models as graphic interfaces. The case studies presented belong to the wide repertoire of mosaics of Ravenna. One of the most peculiar characteristics of mosaics that often limits their digital survey is their multi-scale complexity; nevertheless their models could be used in 3d information systems, for digital exhibitions, for reconstruction aims and to document their conservation conditions in order to conduct restoration interventions in digital environments aiming at speeding and performing more reliable evaluations.

  6. TEHNOLOGIJE 3D TISKALNIKOV

    OpenAIRE

    Kolar, Nataša

    2016-01-01

    Diplomsko delo predstavi razvoj tiskanja skozi čas. Podrobneje so opisani 3D tiskalniki, ki uporabljajo različne tehnologije 3D tiskanja. Predstavljene so različne tehnologije 3D tiskanja, njihova uporaba in narejeni prototipi oz. končni izdelki. Diplomsko delo opiše celoten postopek, od zamisli, priprave podatkov in tiskalnika do izdelave prototipa oz. končnega izdelka.

  7. Imaging a Sustainable Future in 3D

    Science.gov (United States)

    Schuhr, W.; Lee, J. D.; Kanngieser, E.

    2012-07-01

    It is the intention of this paper, to contribute to a sustainable future by providing objective object information based on 3D photography as well as promoting 3D photography not only for scientists, but also for amateurs. Due to the presentation of this article by CIPA Task Group 3 on "3D Photographs in Cultural Heritage", the presented samples are masterpieces of historic as well as of current 3D photography concentrating on cultural heritage. In addition to a report on exemplarily access to international archives of 3D photographs, samples for new 3D photographs taken with modern 3D cameras, as well as by means of a ground based high resolution XLITE staff camera and also 3D photographs taken from a captive balloon and the use of civil drone platforms are dealt with. To advise on optimum suited 3D methodology, as well as to catch new trends in 3D, an updated synoptic overview of the 3D visualization technology, even claiming completeness, has been carried out as a result of a systematic survey. In this respect, e.g., today's lasered crystals might be "early bird" products in 3D, which, due to lack in resolution, contrast and color, remember to the stage of the invention of photography.

  8. Diagnostic Value and Surgical Implications of the 3D DW-SSFP MRI On the Management of Patients with Brachial Plexus Injuries

    Science.gov (United States)

    Qin, Ben-Gang; Yang, Jian-Tao; Yang, Yi; Wang, Hong-Gang; Fu, Guo; Gu, Li-Qiang; Li, Ping; Zhu, Qing-Tang; Liu, Xiao-Lin; Zhu, Jia-Kai

    2016-01-01

    Three-dimensional diffusion-weighted steady-state free precession (3D DW-SSFP) of high-resolution magnetic resonance has emerged as a promising method to visualize the peripheral nerves. In this study, the application value of 3D DW-SSFP brachial plexus imaging in the diagnosis of brachial plexus injury (BPI) was investigated. 33 patients with BPI were prospectively examined using 3D DW-SSFP MR neurography (MRN) of brachial plexus. Results of 3D DW-SSFP MRN were compared with intraoperative findings and measurements of electromyogram (EMG) or somatosensory evoked potentials (SEP) for each injured nerve root. 3D DW-SSFP MRN of brachial plexus has enabled good visualization of the small components of the brachial plexus. The postganglionic section of the brachial plexus was clearly visible in 26 patients, while the preganglionic section of the brachial plexus was clearly visible in 22 patients. Pseudomeningoceles were commonly observed in 23 patients. Others finding of MRN of brachial plexus included spinal cord offset (in 16 patients) and spinal cord deformation (in 6 patients). As for the 3D DW-SSFP MRN diagnosis of preganglionic BPI, the sensitivity, the specificity and the accuracy were respectively 96.8%, 90.29%, and 94.18%. 3D DW-SSFP MRN of brachial plexus improve visualization of brachial plexus and benefit to determine the extent of injury. PMID:27782162

  9. Do TQM interventions change management culture? Findings and implications.

    Science.gov (United States)

    Gerowitz, M B

    1998-01-01

    This study assesses the impact of TQM/CQI interventions on the culture and performance of top management teams. The findings suggest culture is related to performance but that TQM/CQI interventions are not associated with either performance or culture change. Implications for additional research and for practice are discussed.

  10. 3D virtuel udstilling

    DEFF Research Database (Denmark)

    Tournay, Bruno; Rüdiger, Bjarne

    2006-01-01

    3d digital model af Arkitektskolens gård med virtuel udstilling af afgangsprojekter fra afgangen sommer 2006. 10 s.......3d digital model af Arkitektskolens gård med virtuel udstilling af afgangsprojekter fra afgangen sommer 2006. 10 s....

  11. Human in vitro 3D co-culture model to engineer vascularized bone-mimicking tissues combining computational tools and statistical experimental approach.

    Science.gov (United States)

    Bersini, Simone; Gilardi, Mara; Arrigoni, Chiara; Talò, Giuseppe; Zamai, Moreno; Zagra, Luigi; Caiolfa, Valeria; Moretti, Matteo

    2016-01-01

    The generation of functional, vascularized tissues is a key challenge for both tissue engineering applications and the development of advanced in vitro models analyzing interactions among circulating cells, endothelium and organ-specific microenvironments. Since vascularization is a complex process guided by multiple synergic factors, it is critical to analyze the specific role that different experimental parameters play in the generation of physiological tissues. Our goals were to design a novel meso-scale model bridging the gap between microfluidic and macro-scale studies, and high-throughput screen the effects of multiple variables on the vascularization of bone-mimicking tissues. We investigated the influence of endothelial cell (EC) density (3-5 Mcells/ml), cell ratio among ECs, mesenchymal stem cells (MSCs) and osteo-differentiated MSCs (1:1:0, 10:1:0, 10:1:1), culture medium (endothelial, endothelial + angiopoietin-1, 1:1 endothelial/osteo), hydrogel type (100%fibrin, 60%fibrin+40%collagen), tissue geometry (2 × 2 × 2, 2 × 2 × 5 mm(3)). We optimized the geometry and oxygen gradient inside hydrogels through computational simulations and we analyzed microvascular network features including total network length/area and vascular branch number/length. Particularly, we employed the "Design of Experiment" statistical approach to identify key differences among experimental conditions. We combined the generation of 3D functional tissue units with the fine control over the local microenvironment (e.g. oxygen gradients), and developed an effective strategy to enable the high-throughput screening of multiple experimental parameters. Our approach allowed to identify synergic correlations among critical parameters driving microvascular network development within a bone-mimicking environment and could be translated to any vascularized tissue. Copyright © 2015 Elsevier Ltd. All rights reserved.

  12. Structure Segmentation and Transfer Faults in the Marcellus Shale, Clearfield County, Pennsylvania: Implications for Gas Recovery Efficiency and Risk Assessment Using 3D Seismic Attribute Analysis

    Science.gov (United States)

    Roberts, Emily D.

    The Marcellus Shale has become an important unconventional gas reservoir in the oil and gas industry. Fractures within this organic-rich black shale serve as an important component of porosity and permeability useful in enhancing production. Horizontal drilling is the primary approach for extracting hydrocarbons in the Marcellus Shale. Typically, wells are drilled perpendicular to natural fractures in an attempt to intersect fractures for effective hydraulic stimulation. If the fractures are contained within the shale, then hydraulic fracturing can enhance permeability by further breaking the already weakened rock. However, natural fractures can affect hydraulic stimulations by absorbing and/or redirecting the energy away from the wellbore, causing a decreased efficiency in gas recovery, as has been the case for the Clearfield County, Pennsylvania study area. Estimating appropriate distances away from faults and fractures, which may limit hydrocarbon recovery, is essential to reducing the risk of injection fluid migration along these faults. In an attempt to mitigate the negative influences of natural fractures on hydrocarbon extraction within the Marcellus Shale, fractures were analyzed through the aid of both traditional and advanced seismic attributes including variance, curvature, ant tracking, and waveform model regression. Through the integration of well log interpretations and seismic data, a detailed assessment of structural discontinuities that may decrease the recovery efficiency of hydrocarbons was conducted. High-quality 3D seismic data in Central Pennsylvania show regional folds and thrusts above the major detachment interval of the Salina Salt. In addition to the regional detachment folds and thrusts, cross-regional, northwest-trending lineaments were mapped. These lineaments may pose a threat to hydrocarbon productivity and recovery efficiency due to faults and fractures acting as paths of least resistance for induced hydraulic stimulation fluids

  13. Blender 3D cookbook

    CERN Document Server

    Valenza, Enrico

    2015-01-01

    This book is aimed at the professionals that already have good 3D CGI experience with commercial packages and have now decided to try the open source Blender and want to experiment with something more complex than the average tutorials on the web. However, it's also aimed at the intermediate Blender users who simply want to go some steps further.It's taken for granted that you already know how to move inside the Blender interface, that you already have 3D modeling knowledge, and also that of basic 3D modeling and rendering concepts, for example, edge-loops, n-gons, or samples. In any case, it'

  14. Globalisation And African Cultural Heritage Erosion: Implications For ...

    African Journals Online (AJOL)

    Globalisation And African Cultural Heritage Erosion: Implications For Policy. ... Moreover, economic growth and development have frequently been ... and technology for sustainable development and local community livelihoods. Keywords: ...

  15. 3D Digital Modelling

    DEFF Research Database (Denmark)

    Hundebøl, Jesper

    wave of new building information modelling tools demands further investigation, not least because of industry representatives' somewhat coarse parlance: Now the word is spreading -3D digital modelling is nothing less than a revolution, a shift of paradigm, a new alphabet... Research qeustions. Based...... on empirical probes (interviews, observations, written inscriptions) within the Danish construction industry this paper explores the organizational and managerial dynamics of 3D Digital Modelling. The paper intends to - Illustrate how the network of (non-)human actors engaged in the promotion (and arrest) of 3......D Modelling (in Denmark) stabilizes - Examine how 3D Modelling manifests itself in the early design phases of a construction project with a view to discuss the effects hereof for i.a. the management of the building process. Structure. The paper introduces a few, basic methodological concepts...

  16. DELTA 3D PRINTER

    Directory of Open Access Journals (Sweden)

    ȘOVĂILĂ Florin

    2016-07-01

    Full Text Available 3D printing is a very used process in industry, the generic name being “rapid prototyping”. The essential advantage of a 3D printer is that it allows the designers to produce a prototype in a very short time, which is tested and quickly remodeled, considerably reducing the required time to get from the prototype phase to the final product. At the same time, through this technique we can achieve components with very precise forms, complex pieces that, through classical methods, could have been accomplished only in a large amount of time. In this paper, there are presented the stages of a 3D model execution, also the physical achievement after of a Delta 3D printer after the model.

  17. Professional Papervision3D

    CERN Document Server

    Lively, Michael

    2010-01-01

    Professional Papervision3D describes how Papervision3D works and how real world applications are built, with a clear look at essential topics such as building websites and games, creating virtual tours, and Adobe's Flash 10. Readers learn important techniques through hands-on applications, and build on those skills as the book progresses. The companion website contains all code examples, video step-by-step explanations, and a collada repository.

  18. AE3D

    Energy Technology Data Exchange (ETDEWEB)

    2016-06-20

    AE3D solves for the shear Alfven eigenmodes and eigenfrequencies in a torodal magnetic fusion confinement device. The configuration can be either 2D (e.g. tokamak, reversed field pinch) or 3D (e.g. stellarator, helical reversed field pinch, tokamak with ripple). The equations solved are based on a reduced MHD model and sound wave coupling effects are not currently included.

  19. Implication of the development of 3D printing technology for education%三维打印技术的发展对教育的启迪

    Institute of Scientific and Technical Information of China (English)

    黎加厚; 杨晓哲; 杨高云

    2013-01-01

    在介绍三维打印技术现状及其未来发展趋势的基础上,进一步具体介绍了在上海市和田路小学开展运用3D打印机培养学生创新思维的实验.和田路小学开展利用三维打印技术的校本课程实践表明,让学生从小就接触、体验、掌握新兴的三维打印技术,让学生体验未来工业生产的新模式,有利于提升学生的想象力和创造力.%The paper first introduces the actuality of 3D printing technology and its future development trend, and then further introduces the experimental use of 3D printer to cultivate- the innovative thinking of students in Shanghai Hetian Road Primary School. The practice of school-based class of using 3D printing technology in Hetian Road Primary School shows that letting students contact, experience and grasp the emerging 3D printing technology and experience the new model of future industrial production at young age will help improve students'imagination and creativity.

  20. Interactive 3D Hybrid PET/CT Imaging in the Identification of Myocardial Viability in Patients After Myocardial Infarction: Feasibility Study and Clinical Implications

    Directory of Open Access Journals (Sweden)

    Yen-Wen Wu

    2008-06-01

    Conclusion: We present a protocol to acquire CT coronary angiography and PET data and to visualize 3D fused images with an interactive visualization interface. This image coregistration is potentially useful to facilitate the process of image interpretation and decision-making.

  1. Magnetotelluric Transfer Functions: Phase Tensor and Tipper Vector above a Simple Anisotropic Three-Dimensional Conductivity Anomaly and Implications for 3D Isotropic Inversion

    Science.gov (United States)

    Löwer, Alexander; Junge, Andreas

    2017-05-01

    The influence of anisotropic conductivity structures on magnetotelluric transfer functions is not easy to analyse in its entire complexity. In this study, we investigate the spatial and frequency-dependent behaviour of phase tensors and tipper vectors above a 3D anisotropic conductivity anomaly. The anomaly consists of a simple cubic block embedded in a homogeneous half space. Using a 3D FD code, we compare an isotropic, 2 anisotropic models with an anisotropy factor of 10 and one anisotropic model with the anisotropy factor of 100. The results show characteristic differences between the isotropic and anisotropic cases. For the anisotropic anomalies, the tipper vectors are parallel over the entire area despite the 3D geometry of the anomalous body. The size of the tipper vectors depends on the position of the site relative to the anomaly's boundaries and the direction of the anisotropic strike. Above the anomalous anisotropic body, the main diagonal elements of the phase tensor show the well-known split. Outside the anomaly, the phase tensor principal axis rotates according to the site position in contrast to the constant tipper direction. The 3D inversion of the forward data using an isotropic 3D code (ModEM) yields a very good fit for all cases. Whereas the inversion result matches the isotropic model, wave-like structures with high conductivity contrast occur for the anisotropic models. These structures extend far beyond the extension of the original anomalous body. Thus, the study reveals important indications of the existence of anisotropic conductivity structures for observed magnetotelluric transfer functions.

  2. International and Cultural Implications on Internationalization Analysis of Multinational Firms

    Directory of Open Access Journals (Sweden)

    José G. Vargas-Hernández

    2015-08-01

    Full Text Available This paper is aims to analyze some of the institutional and cultural implications on internationalization analysis of multinational firms. The analysis begins questioning what the main institutional and cultural variables are considered in the involvement of internationalization of multinational firms. To answer this question, a literature review types approach in areas like internationalization of multinational firms based on institutional and cultural frameworks is followed. Secondly, these institutional and cultural variables are analyzed to integrate findings. Finally, the paper argues the need to design a better institutional and cultural balance among the development of a glocal-regional transformation, convergence and governance.

  3. 3D Projection Installations

    DEFF Research Database (Denmark)

    Halskov, Kim; Johansen, Stine Liv; Bach Mikkelsen, Michelle

    2014-01-01

    Three-dimensional projection installations are particular kinds of augmented spaces in which a digital 3-D model is projected onto a physical three-dimensional object, thereby fusing the digital content and the physical object. Based on interaction design research and media studies, this article...... contributes to the understanding of the distinctive characteristics of such a new medium, and identifies three strategies for designing 3-D projection installations: establishing space; interplay between the digital and the physical; and transformation of materiality. The principal empirical case, From...... Fingerplan to Loop City, is a 3-D projection installation presenting the history and future of city planning for the Copenhagen area in Denmark. The installation was presented as part of the 12th Architecture Biennale in Venice in 2010....

  4. 3D Spectroscopic Instrumentation

    CERN Document Server

    Bershady, Matthew A

    2009-01-01

    In this Chapter we review the challenges of, and opportunities for, 3D spectroscopy, and how these have lead to new and different approaches to sampling astronomical information. We describe and categorize existing instruments on 4m and 10m telescopes. Our primary focus is on grating-dispersed spectrographs. We discuss how to optimize dispersive elements, such as VPH gratings, to achieve adequate spectral resolution, high throughput, and efficient data packing to maximize spatial sampling for 3D spectroscopy. We review and compare the various coupling methods that make these spectrographs ``3D,'' including fibers, lenslets, slicers, and filtered multi-slits. We also describe Fabry-Perot and spatial-heterodyne interferometers, pointing out their advantages as field-widened systems relative to conventional, grating-dispersed spectrographs. We explore the parameter space all these instruments sample, highlighting regimes open for exploitation. Present instruments provide a foil for future development. We give an...

  5. EDUCATIONAL IMPLICATIONS OF CONTACT BETWEEN CULTURES

    Directory of Open Access Journals (Sweden)

    Irene Verde Peleato

    2010-11-01

    Full Text Available This article responds to the interest that continues to raise the meeting of cultures present in school and all that involves in the teaching task. In particular, from an investigation conducted on bilingual education in California, raised some thoughts that should be taken into account when working in school settings of cultural and linguistic diversity.

  6. Cultural Diversity: Implications For Workplace Management

    Directory of Open Access Journals (Sweden)

    Donatus I. Amaram

    2011-07-01

    Full Text Available The acceptance and management of cultural diversity have been promoted and touted as a positive tool in social and organizational engineering aimed at solving and preventing group dynamics problems in both business organizations and society as well. Positive attributes of cultural integration in business organizations have received fair and significant attention in the past two decades. What have not been sufficiently presented are the challenges and pitfalls inherent in the management of culturally diverse work groups. For the practicing manager, there is a need to know when and where mono- and multi-cultural arrangements may be preferred. This paper reviews relevant research findings that can be used for building effective paradigms in the management of cultural diversity in the workplace.

  7. Simple and Versatile Turbidimetric Monitoring of Bacterial Growth in Liquid Cultures Using a Customized 3D Printed Culture Tube Holder and a Miniaturized Spectrophotometer: Application to Facultative and Strictly Anaerobic Bacteria

    Science.gov (United States)

    Maia, Margarida R. G.; Marques, Sara; Cabrita, Ana R. J.; Wallace, R. John; Thompson, Gertrude; Fonseca, António J. M.; Oliveira, Hugo M.

    2016-01-01

    Here we introduce a novel strategy for turbidimetric monitoring of bacterial growth in liquid culture. The instrumentation comprises a light source, a customized 3D printed culture tube holder and a miniaturized spectrophotometer, connected through optical cables. Due to its small footprint and the possibility to operate with external light, bacterial growth was directly monitored from culture tubes in a simple and versatile fashion. This new portable measurement technique was used to monitor the growth of facultative (Escherichia coli ATCC/25922, and Staphylococcus aureus ATCC/29213) and strictly (Butyrivibrio fibrisolvens JW11, Butyrivibrio proteoclasticus P18, and Propionibacterium acnes DSMZ 1897) anaerobic bacteria. For E. coli and S. aureus, the growth rates calculated from normalized optical density values were compared with those ones obtained using a benchtop spectrophotometer without significant differences (P = 0.256). For the strictly anaerobic species, a high precision (relative standard deviation < 3.5%) was observed between replicates up to 48 h. Regarding its potential for customization, this manifold could accommodate further developments for customized turbidimetric monitoring, such as the use of light-emitting diodes as a light source or flow cells. PMID:27630632

  8. Simple and versatile turbidimetric monitoring of bacterial growth in liquid cultures using a customized 3D printed culture tube holder and a miniaturized spectrophotometer: application to facultative and strictly anaerobic bacteria

    Directory of Open Access Journals (Sweden)

    Margarida R. G. Maia

    2016-08-01

    Full Text Available Here we introduce a novel strategy for turbidimetric monitoring of bacterial growth in liquid culture. The instrumentation comprises a light source, a customized 3D printed culture tube holder and a miniaturized spectrophotometer, connected through optical cables. Due to its small footprint and the possibility to operate with external light, bacterial growth was directly monitored from culture tubes in a simple and versatile fashion. This new portable measurement technique was used to monitor the growth of facultative (Escherichia coli ATCC/25922, and Staphylococcus aureus ATCC/29213 and strictly (Butyrivibrio fibrisolvens JW11, Butyrivibrio proteoclasticus P18, and Propionibacterium acnes DSMZ 1897 anaerobic bacteria. For E. coli and S. aureus, the growth rates calculated from normalized optical density values were compared with those ones obtained using a benchtop spectrophotometer without significant differences (P = 0.256. For the strictly anaerobic species, a high precision (RSD < 3.5% was observed between replicates up to 48 h. Regarding its potential for customization, this manifold could accommodate further developments for customized turbidimetric monitoring, such as the use of light-emitting diodes as a light source or flow cells.

  9. Simple and Versatile Turbidimetric Monitoring of Bacterial Growth in Liquid Cultures Using a Customized 3D Printed Culture Tube Holder and a Miniaturized Spectrophotometer: Application to Facultative and Strictly Anaerobic Bacteria.

    Science.gov (United States)

    Maia, Margarida R G; Marques, Sara; Cabrita, Ana R J; Wallace, R John; Thompson, Gertrude; Fonseca, António J M; Oliveira, Hugo M

    2016-01-01

    Here we introduce a novel strategy for turbidimetric monitoring of bacterial growth in liquid culture. The instrumentation comprises a light source, a customized 3D printed culture tube holder and a miniaturized spectrophotometer, connected through optical cables. Due to its small footprint and the possibility to operate with external light, bacterial growth was directly monitored from culture tubes in a simple and versatile fashion. This new portable measurement technique was used to monitor the growth of facultative (Escherichia coli ATCC/25922, and Staphylococcus aureus ATCC/29213) and strictly (Butyrivibrio fibrisolvens JW11, Butyrivibrio proteoclasticus P18, and Propionibacterium acnes DSMZ 1897) anaerobic bacteria. For E. coli and S. aureus, the growth rates calculated from normalized optical density values were compared with those ones obtained using a benchtop spectrophotometer without significant differences (P = 0.256). For the strictly anaerobic species, a high precision (relative standard deviation < 3.5%) was observed between replicates up to 48 h. Regarding its potential for customization, this manifold could accommodate further developments for customized turbidimetric monitoring, such as the use of light-emitting diodes as a light source or flow cells.

  10. Interaktiv 3D design

    DEFF Research Database (Denmark)

    Villaume, René Domine; Ørstrup, Finn Rude

    2002-01-01

    Projektet undersøger potentialet for interaktiv 3D design via Internettet. Arkitekt Jørn Utzons projekt til Espansiva blev udviklet som et byggesystem med det mål, at kunne skabe mangfoldige planmuligheder og mangfoldige facade- og rumudformninger. Systemets bygningskomponenter er digitaliseret som...... 3D elementer og gjort tilgængelige. Via Internettet er det nu muligt at sammenstille og afprøve en uendelig  række bygningstyper som  systemet blev tænkt og udviklet til....

  11. Cross-cultural discussions in a 3D virtual environment and their affordances for learners’ motivation and foreign language discussion skills

    NARCIS (Netherlands)

    Jauregi Ondarra, M.K.; Kuure, L.; Bastian, P.; Reinhardt, D.; Koivisto, T.

    2015-01-01

    Within the European TILA project a case study was carried out where pupils from schools in Finland and the Netherlands engaged in debating sessions using the 3D virtual world of OpenSim once a week in a period of 5 weeks. The case study had two main objectives: (1) to study the impact that the discu

  12. Cross-cultural discussions in a 3D virtual environment and their affordances for learners’ motivation and foreign language discussion skills

    NARCIS (Netherlands)

    Jauregi Ondarra, M.K.|info:eu-repo/dai/nl/276318102; Kuure, L.; Bastian, P.; Reinhardt, D.; Koivisto, T.

    2015-01-01

    Within the European TILA project a case study was carried out where pupils from schools in Finland and the Netherlands engaged in debating sessions using the 3D virtual world of OpenSim once a week in a period of 5 weeks. The case study had two main objectives: (1) to study the impact that the

  13. The Idaho Virtualization Laboratory 3D Pipeline

    Directory of Open Access Journals (Sweden)

    Nicholas A. Holmer

    2014-05-01

    Full Text Available Three dimensional (3D virtualization and visualization is an important component of industry, art, museum curation and cultural heritage, yet the step by step process of 3D virtualization has been little discussed. Here we review the Idaho Virtualization Laboratory’s (IVL process of virtualizing a cultural heritage item (artifact from start to finish. Each step is thoroughly explained and illustrated including how the object and its metadata are digitally preserved and ultimately distributed to the world.

  14. 3D GEO: AN ALTERNATIVE APPROACH

    OpenAIRE

    2016-01-01

    The expression GEO is mostly used to denote relation to the earth. However it should not be confined to what is related to the earth's surface, as other objects also need three dimensional representation and documentation, like cultural heritage objects. They include both tangible and intangible ones. In this paper the 3D data acquisition and 3D modelling of cultural heritage assets are briefly described and their significance is also highlighted. Moreover the organization of such information...

  15. Use of INSAT-3D sounder and imager radiances in the 4D-VAR data assimilation system and its implications in the analyses and forecasts

    Science.gov (United States)

    Indira Rani, S.; Taylor, Ruth; George, John P.; Rajagopal, E. N.

    2016-05-01

    INSAT-3D, the first Indian geostationary satellite with sounding capability, provides valuable information over India and the surrounding oceanic regions which are pivotal to Numerical Weather Prediction. In collaboration with UK Met Office, NCMRWF developed the assimilation capability of INSAT-3D Clear Sky Brightness Temperature (CSBT), both from the sounder and imager, in the 4D-Var assimilation system being used at NCMRWF. Out of the 18 sounder channels, radiances from 9 channels are selected for assimilation depending on relevance of the information in each channel. The first three high peaking channels, the CO2 absorption channels and the three water vapor channels (channel no. 10, 11, and 12) are assimilated both over land and Ocean, whereas the window channels (channel no. 6, 7, and 8) are assimilated only over the Ocean. Measured satellite radiances are compared with that from short range forecasts to monitor the data quality. This is based on the assumption that the observed satellite radiances are free from calibration errors and the short range forecast provided by NWP model is free from systematic errors. Innovations (Observation - Forecast) before and after the bias correction are indicative of how well the bias correction works. Since the biases vary with air-masses, time, scan angle and also due to instrument degradation, an accurate bias correction algorithm for the assimilation of INSAT-3D sounder radiance is important. This paper discusses the bias correction methods and other quality controls used for the selected INSAT-3D sounder channels and the impact of bias corrected radiance in the data assimilation system particularly over India and surrounding oceanic regions.

  16. 3D Wire 2015

    DEFF Research Database (Denmark)

    Jordi, Moréton; F, Escribano; J. L., Farias

    This document is a general report on the implementation of gamification in 3D Wire 2015 event. As the second gamification experience in this event, we have delved deeply in the previous objectives (attracting public areas less frequented exhibition in previous years and enhance networking) and ha......, improves socialization and networking, improves media impact, improves fun factor and improves encouragement of the production team....

  17. Shaping 3-D boxes

    DEFF Research Database (Denmark)

    Stenholt, Rasmus; Madsen, Claus B.

    2011-01-01

    Enabling users to shape 3-D boxes in immersive virtual environments is a non-trivial problem. In this paper, a new family of techniques for creating rectangular boxes of arbitrary position, orientation, and size is presented and evaluated. These new techniques are based solely on position data...

  18. Tangible 3D Modelling

    DEFF Research Database (Denmark)

    Hejlesen, Aske K.; Ovesen, Nis

    2012-01-01

    This paper presents an experimental approach to teaching 3D modelling techniques in an Industrial Design programme. The approach includes the use of tangible free form models as tools for improving the overall learning. The paper is based on lecturer and student experiences obtained through facil...

  19. 3D Geometric Modeling of the Abu Madi Reservoirs and Its Implication on the Gas Development in Baltim Area (Offshore Nile Delta, Egypt

    Directory of Open Access Journals (Sweden)

    Mohamed I. Abdel-Fattah

    2015-01-01

    Full Text Available 3D geometric modeling has received renewed attention recently, in the context of visual scene understanding. The reservoir geometry of the Baltim fields is described by significant elements, such as thickness, depth maps, and fault planes, resulting from an interpretation based on seismic and well data. Uncertainties affect these elements throughout the entire interpretation process. They have some bearing on the geometric shape and subsequently on the gross reservoir volume (GRV of the fields. This uncertainty on GRV also impacts volumes of hydrocarbons in place, reserves, and production profiles. Thus, the assessment of geometrical uncertainties is an essential first step in a field study for evaluation, development, and optimization purposes. Seismic data are best integrated with well and reservoir information. A 3D geometric model of the Late Messinian Abu Madi reservoirs in the time and depth domain is used to investigate the influence of the reservoir geometry on the gas entrapment. Important conceptual conclusions about the reservoir system behavior are obtained using this model. The results show that the reservoir shape influences the seismic response of the incised Abu Madi Paleovalley, making it necessary to account for 3D effects in order to obtain accurate results.

  20. The geological implications of some 3-D 'Fishnet' seismic images of Guelph pinnacle reefs in Ontario : a different perspective on their growth, destruction and complexity

    Energy Technology Data Exchange (ETDEWEB)

    Bailey, S.M.B. [Bailey Geological Services Ltd., London, ON (Canada)

    2002-07-01

    Exploration of underground oil and gas reservoirs has been enhanced since the development of 3-D seismic technology, which provides the ability to view structural anomalies, salt domes or lithologic buildups. 3-D seismic images were used to view the true geometry of carbonate structures within Silurian Guelph reefs in southwestern Ontario. The images provided a better understanding of how they grew, plus when and how they were destroyed by post-depositional weathering processes. 3-D seismic images allow oil and gas companies to explore and develop a series of wells through tight and porous segments of a reservoir with extreme precision. The Guelph reefs of Ontario are Middle Silurian in age and are part of an entire complex of reefs, banks, barriers and associated facies which grew in a circular fashion around a structural basement sag known as the Michigan Basin. The author explains how two reefs of the same physical height can be so evidently different in physical appearance. He presents his interpretation of how the Bentpath East Pinnacle Reef and the Tipperary Pinnacle reef underwent completely different histories of weathering, destruction, and construction. 16 refs., 1 tab., 33 figs.

  1. Avenues of Influence: Cross-Cultural Implications

    Science.gov (United States)

    2014-04-01

    adapt the United Nations (UN taxonomy of 23 culture groups to four cultural groups based on NATO School 2008 (or 2009, when available) matriculation ...ungegndivisions.htm#ecsee). Minor adjustments to the UN taxonomy were proposed, as some countries on the NATO School 2008 matriculation were listed in...Dialogue, and other nations. Of NATO School students matriculating in 2008, just 17% were from native-English speaking countries. Of the

  2. 3D photoacoustic imaging

    Science.gov (United States)

    Carson, Jeffrey J. L.; Roumeliotis, Michael; Chaudhary, Govind; Stodilka, Robert Z.; Anastasio, Mark A.

    2010-06-01

    Our group has concentrated on development of a 3D photoacoustic imaging system for biomedical imaging research. The technology employs a sparse parallel detection scheme and specialized reconstruction software to obtain 3D optical images using a single laser pulse. With the technology we have been able to capture 3D movies of translating point targets and rotating line targets. The current limitation of our 3D photoacoustic imaging approach is its inability ability to reconstruct complex objects in the field of view. This is primarily due to the relatively small number of projections used to reconstruct objects. However, in many photoacoustic imaging situations, only a few objects may be present in the field of view and these objects may have very high contrast compared to background. That is, the objects have sparse properties. Therefore, our work had two objectives: (i) to utilize mathematical tools to evaluate 3D photoacoustic imaging performance, and (ii) to test image reconstruction algorithms that prefer sparseness in the reconstructed images. Our approach was to utilize singular value decomposition techniques to study the imaging operator of the system and evaluate the complexity of objects that could potentially be reconstructed. We also compared the performance of two image reconstruction algorithms (algebraic reconstruction and l1-norm techniques) at reconstructing objects of increasing sparseness. We observed that for a 15-element detection scheme, the number of measureable singular vectors representative of the imaging operator was consistent with the demonstrated ability to reconstruct point and line targets in the field of view. We also observed that the l1-norm reconstruction technique, which is known to prefer sparseness in reconstructed images, was superior to the algebraic reconstruction technique. Based on these findings, we concluded (i) that singular value decomposition of the imaging operator provides valuable insight into the capabilities of

  3. Unoriented 3d TFTs

    CERN Document Server

    Bhardwaj, Lakshya

    2016-01-01

    This paper generalizes two facts about oriented 3d TFTs to the unoriented case. On one hand, it is known that oriented 3d TFTs having a topological boundary condition admit a state-sum construction known as the Turaev-Viro construction. This is related to the string-net construction of fermionic phases of matter. We show how Turaev-Viro construction can be generalized to unoriented 3d TFTs. On the other hand, it is known that the "fermionic" versions of oriented TFTs, known as Spin-TFTs, can be constructed in terms of "shadow" TFTs which are ordinary oriented TFTs with an anomalous Z_2 1-form symmetry. We generalize this correspondence to Pin+ TFTs by showing that they can be constructed in terms of ordinary unoriented TFTs with anomalous Z_2 1-form symmetry having a mixed anomaly with time-reversal symmetry. The corresponding Pin+ TFT does not have any anomaly for time-reversal symmetry however and hence it can be unambiguously defined on a non-orientable manifold. In case a Pin+ TFT admits a topological bou...

  4. C-peptide and zinc delivery to erythrocytes requires the presence of albumin: implications in diabetes explored with a 3D-printed fluidic device.

    Science.gov (United States)

    Liu, Yueli; Chen, Chengpeng; Summers, Suzanne; Medawala, Wathsala; Spence, Dana M

    2015-05-01

    People with type 1 diabetes (T1D) must administer insulin exogenously due to the destruction of their pancreatic β-cells. Endogenous insulin is stored in β-cell granules along with C-peptide, a 31 amino acid peptide that is secreted from these granules in amounts equal to insulin. Exogenous co-administration of C-peptide with insulin has proven to reduce diabetes-associated complications in animals and humans. The exact mechanism of C-peptide's beneficial effects after secretion from the β-cell granules is not completely understood, thus hindering its development as an exogenously administered hormone. Monitoring tissue-to-tissue communication using a 3D-printed microfluidic device revealed that zinc and C-peptide are being delivered to erythrocytes by albumin. Upon delivery, erythrocyte-derived ATP increased by >50%, as did endothelium-derived NO, which was measured downstream in the 3D-printed device. Our results suggest that hormone replacement therapy in diabetes may be improved by exogenous administration of a C-peptide ensemble that includes zinc and albumin.

  5. Tooteko: a Case Study of Augmented Reality for AN Accessible Cultural Heritage. Digitization, 3d Printing and Sensors for AN Audio-Tactile Experience

    Science.gov (United States)

    D'Agnano, F.; Balletti, C.; Guerra, F.; Vernier, P.

    2015-02-01

    Tooteko is a smart ring that allows to navigate any 3D surface with your finger tips and get in return an audio content that is relevant in relation to the part of the surface you are touching in that moment. Tooteko can be applied to any tactile surface, object or sheet. However, in a more specific domain, it wants to make traditional art venues accessible to the blind, while providing support to the reading of the work for all through the recovery of the tactile dimension in order to facilitate the experience of contact with art that is not only "under glass." The system is made of three elements: a high-tech ring, a tactile surface tagged with NFC sensors, and an app for tablet or smartphone. The ring detects and reads the NFC tags and, thanks to the Tooteko app, communicates in wireless mode with the smart device. During the tactile navigation of the surface, when the finger reaches a hotspot, the ring identifies the NFC tag and activates, through the app, the audio track that is related to that specific hotspot. Thus a relevant audio content relates to each hotspot. The production process of the tactile surfaces involves scanning, digitization of data and 3D printing. The first experiment was modelled on the facade of the church of San Michele in Isola, made by Mauro Codussi in the late fifteenth century, and which marks the beginning of the Renaissance in Venice. Due to the absence of recent documentation on the church, the Correr Museum asked the Laboratorio di Fotogrammetria to provide it with the aim of setting up an exhibition about the order of the Camaldolesi, owners of the San Michele island and church. The Laboratorio has made the survey of the facade through laser scanning and UAV photogrammetry. The point clouds were the starting point for prototypation and 3D printing on different supports. The idea of the integration between a 3D printed tactile surface and sensors was born as a final thesis project at the Postgraduate Mastercourse in Digital

  6. Three-dimensional (3D) printing of mouse primary hepatocytes to generate 3D hepatic structure

    Science.gov (United States)

    Kim, Yohan; Kang, Kyojin; Jeong, Jaemin; Paik, Seung Sam; Kim, Ji Sook; Park, Su A; Kim, Wan Doo; Park, Jisun

    2017-01-01

    Purpose The major problem in producing artificial livers is that primary hepatocytes cannot be cultured for many days. Recently, 3-dimensional (3D) printing technology draws attention and this technology regarded as a useful tool for current cell biology. By using the 3D bio-printing, these problems can be resolved. Methods To generate 3D bio-printed structures (25 mm × 25 mm), cells-alginate constructs were fabricated by 3D bio-printing system. Mouse primary hepatocytes were isolated from the livers of 6–8 weeks old mice by a 2-step collagenase method. Samples of 4 × 107 hepatocytes with 80%–90% viability were printed with 3% alginate solution, and cultured with well-defined culture medium for primary hepatocytes. To confirm functional ability of hepatocytes cultured on 3D alginate scaffold, we conducted quantitative real-time polymerase chain reaction and immunofluorescence with hepatic marker genes. Results Isolated primary hepatocytes were printed with alginate. The 3D printed hepatocytes remained alive for 14 days. Gene expression levels of Albumin, HNF-4α and Foxa3 were gradually increased in the 3D structures. Immunofluorescence analysis showed that the primary hepatocytes produced hepatic-specific proteins over the same period of time. Conclusion Our research indicates that 3D bio-printing technique can be used for long-term culture of primary hepatocytes. It can therefore be used for drug screening and as a potential method of producing artificial livers. PMID:28203553

  7. Three-dimensional (3D) printing of mouse primary hepatocytes to generate 3D hepatic structure.

    Science.gov (United States)

    Kim, Yohan; Kang, Kyojin; Jeong, Jaemin; Paik, Seung Sam; Kim, Ji Sook; Park, Su A; Kim, Wan Doo; Park, Jisun; Choi, Dongho

    2017-02-01

    The major problem in producing artificial livers is that primary hepatocytes cannot be cultured for many days. Recently, 3-dimensional (3D) printing technology draws attention and this technology regarded as a useful tool for current cell biology. By using the 3D bio-printing, these problems can be resolved. To generate 3D bio-printed structures (25 mm × 25 mm), cells-alginate constructs were fabricated by 3D bio-printing system. Mouse primary hepatocytes were isolated from the livers of 6-8 weeks old mice by a 2-step collagenase method. Samples of 4 × 10(7) hepatocytes with 80%-90% viability were printed with 3% alginate solution, and cultured with well-defined culture medium for primary hepatocytes. To confirm functional ability of hepatocytes cultured on 3D alginate scaffold, we conducted quantitative real-time polymerase chain reaction and immunofluorescence with hepatic marker genes. Isolated primary hepatocytes were printed with alginate. The 3D printed hepatocytes remained alive for 14 days. Gene expression levels of Albumin, HNF-4α and Foxa3 were gradually increased in the 3D structures. Immunofluorescence analysis showed that the primary hepatocytes produced hepatic-specific proteins over the same period of time. Our research indicates that 3D bio-printing technique can be used for long-term culture of primary hepatocytes. It can therefore be used for drug screening and as a potential method of producing artificial livers.

  8. Modeling and Accuracy Assessment for 3D-VIRTUAL Reconstruction in Cultural Heritage Using Low-Cost Photogrammetry: Surveying of the "santa MARÍA Azogue" Church's Front

    Science.gov (United States)

    Robleda Prieto, G.; Pérez Ramos, A.

    2015-02-01

    Sometimes it could be difficult to represent "on paper" an architectural idea, a solution, a detail or a newly created element, depending on the complexity what it want be conveyed through its graphical representation but it may be even harder to represent the existing reality. (a building, a detail,...), at least with an acceptable degree of definition and accuracy. As a solution to this hypothetical problem, this paper try to show a methodology to collect measure data by combining different methods or techniques, to obtain the characteristic geometry of architectonic elements, especially in those highly decorated and/or complex geometry, as well as to assess the accuracy of the results obtained, but in an accuracy level enough and not very expensive costs. In addition, we can obtain a 3D recovery model that allows us a strong support, beyond point clouds obtained through another more expensive methods as using laser scanner, to obtain orthoimages. This methodology was used in the study case of the 3D-virtual reconstruction of a main medieval church façade because of the geometrical complexity in many elements as the existing main doorway with archivolts and many details, as well as the rose window located above it so it's inaccessible due to the height.

  9. Geomorphologic characteristics of debris flows in the Ulleung Basin, East Sea (Japan Sea) interpreted from 3-D seismic data and their implications

    Science.gov (United States)

    MO, C.; Park, G.; Lee, G.; Yi, B.; Yoo, D.

    2012-12-01

    We processed and analyzed the 3-D seismic data from the southern central part of the Ulleung Basin, East Sea (Japan Sea) to investigate the geomorphologic characteristics of the debris flows. The data processing included dip moveout, post-stack migration, and acquisition footprint removal. The curvature attributes of the seafloor show numerous bubble- or dot-like features that form a N-S to NNE-SSW trending narrow (ca. 2 km wide) zone in the western part of the area. The bubble-like features correspond to the irregular seafloor in the seismic profiles. At least nine debris flows, which advanced largely north and northeastward, were identified from the seafloor to the sub-seafloor depth of about 300 m. The debris flows are lens- or wedge-shaped in cross section, characterized by structureless or transparent to chaotic internal reflections, and elongate or lobate in plan view. The largest debris flow exceeds the 3D seismic data coverage (16 km by 25 km) and its thickness reaches about 60 m. Some debris flows are very thin and amalgamated or coalesced, making it difficult to interpret the individual flows. The similarity and curvature attributes of the basal contact of some debris flows show numerous long grooves, erosional scars, and bubble- or dot-like features similar to those seen in the seafloor. The grooves, interpreted to be caused by large clasts imbedded at the base of the debris flows, diverge and become slightly wider (decrease in the number of the bubble-like features away from the axis of the debris flows probably suggest decreasing pore fluid pressure toward the edge of the debris flows.

  10. The coupling between idealized 2D eddies and 3D small-scale turbulence in the upper ocean and its implications for oil transport simulations using ENDLESS

    Science.gov (United States)

    Chen, B.; Yang, D.; Meneveau, C. V.; Chamecki, M.

    2016-12-01

    As oil plumes from deep-water blowouts reach the ocean mixed layer (OML), they experience considerable horizontal and vertical dilution due to the action of Langmuir turbulence, submesoscale eddies and Ekman transport. Previous studies using large-eddy simulation (LES) have shown that Langmuir turbulence can impact the transport direction, lateral diffusion and geometry of surface oil plume, depending on the size of oil droplets. However, the large range of relevant length scales, from submesoscale eddies down to small-scale 3D Langmuir turbulence, makes it challenging to accurately reproduce the long-term evolution of oil plumes in the upper ocean using conventional large-eddy simulation strategies. The extended nonperiodic domain large-eddy simulation for scalars (ENDLESS) is a new technique developed as a multi-scale approach to simulate long-term oil plume dispersion at a reasonable computational cost. The basic idea is to simulate Langmuir turbulence on a LES domain with a small but sufficient horizontal domain size to capture the essential physics of the flow field, while simulating the oil plume evolution over an effectively horizontally extended large domain. Moreover, this ENDLESS method permits the superposition of large-scale quasi-2D ocean eddies on the oil advection, allowing for coupling with regional ocean circulation models. Using this new numerical tool, the effects of the interactions between 3D turbulence and 2D eddies on the flow field and on oil dispersion are elucidated by comparing the results from ENDLESS with those from conventional LES. This research was made possible by a grant from The Gulf of Mexico Research Initiative.

  11. 3D and beyond

    Science.gov (United States)

    Fung, Y. C.

    1995-05-01

    This conference on physiology and function covers a wide range of subjects, including the vasculature and blood flow, the flow of gas, water, and blood in the lung, the neurological structure and function, the modeling, and the motion and mechanics of organs. Many technologies are discussed. I believe that the list would include a robotic photographer, to hold the optical equipment in a precisely controlled way to obtain the images for the user. Why are 3D images needed? They are to achieve certain objectives through measurements of some objects. For example, in order to improve performance in sports or beauty of a person, we measure the form, dimensions, appearance, and movements.

  12. Using Cultural Diversity in Teaching Economics: Global Business Implications

    Science.gov (United States)

    Mitry, Darryl J.

    2008-01-01

    Globalization and increasing cross-cultural interactivity have implications for education in general and may also present valuable pedagogical opportunities in the practice of teaching economics for business students. Therefore, the author investigated this proposition and offers some empirical observations from research and teaching experiments.…

  13. Using Cultural Diversity in Teaching Economics: Global Business Implications

    Science.gov (United States)

    Mitry, Darryl J.

    2008-01-01

    Globalization and increasing cross-cultural interactivity have implications for education in general and may also present valuable pedagogical opportunities in the practice of teaching economics for business students. Therefore, the author investigated this proposition and offers some empirical observations from research and teaching experiments.…

  14. 3D Surgical Simulation

    Science.gov (United States)

    Cevidanes, Lucia; Tucker, Scott; Styner, Martin; Kim, Hyungmin; Chapuis, Jonas; Reyes, Mauricio; Proffit, William; Turvey, Timothy; Jaskolka, Michael

    2009-01-01

    This paper discusses the development of methods for computer-aided jaw surgery. Computer-aided jaw surgery allows us to incorporate the high level of precision necessary for transferring virtual plans into the operating room. We also present a complete computer-aided surgery (CAS) system developed in close collaboration with surgeons. Surgery planning and simulation include construction of 3D surface models from Cone-beam CT (CBCT), dynamic cephalometry, semi-automatic mirroring, interactive cutting of bone and bony segment repositioning. A virtual setup can be used to manufacture positioning splints for intra-operative guidance. The system provides further intra-operative assistance with the help of a computer display showing jaw positions and 3D positioning guides updated in real-time during the surgical procedure. The CAS system aids in dealing with complex cases with benefits for the patient, with surgical practice, and for orthodontic finishing. Advanced software tools for diagnosis and treatment planning allow preparation of detailed operative plans, osteotomy repositioning, bone reconstructions, surgical resident training and assessing the difficulties of the surgical procedures prior to the surgery. CAS has the potential to make the elaboration of the surgical plan a more flexible process, increase the level of detail and accuracy of the plan, yield higher operative precision and control, and enhance documentation of cases. Supported by NIDCR DE017727, and DE018962 PMID:20816308

  15. TOWARDS: 3D INTERNET

    Directory of Open Access Journals (Sweden)

    Ms. Swapnali R. Ghadge

    2013-08-01

    Full Text Available In today’s ever-shifting media landscape, it can be a complex task to find effective ways to reach your desired audience. As traditional media such as television continue to lose audience share, one venue in particular stands out for its ability to attract highly motivated audiences and for its tremendous growth potential the 3D Internet. The concept of '3D Internet' has recently come into the spotlight in the R&D arena, catching the attention of many people, and leading to a lot of discussions. Basically, one can look into this matter from a few different perspectives: visualization and representation of information, and creation and transportation of information, among others. All of them still constitute research challenges, as no products or services are yet available or foreseen for the near future. Nevertheless, one can try to envisage the directions that can be taken towards achieving this goal. People who take part in virtual worlds stay online longer with a heightened level of interest. To take advantage of that interest, diverse businesses and organizations have claimed an early stake in this fast-growing market. They include technology leaders such as IBM, Microsoft, and Cisco, companies such as BMW, Toyota, Circuit City, Coca Cola, and Calvin Klein, and scores of universities, including Harvard, Stanford and Penn State.

  16. Implications of Japanese Culture for Cultural Construction of Chinese Agricultural Enterprises

    Institute of Scientific and Technical Information of China (English)

    Ming; XU

    2015-01-01

    The cultural construction in agricultural enterprises in China is not so optimistic especially compared with developed countries. In2015,as the Party Central Committee in China put forward strategy of building a moderately prosperous society in all aspects,it is necessary for Chinese agricultural enterprise to gain implications for cultural construction by studying Japanese culture. Japanese culture features " group spirit,interpersonal relation,absorbing advanced culture and rational spirit" as its quintessence and with " lifetime employment,annual merits,decision making on application,cooperative management ". Chinese agricultural enterprises should find ways for cultural construction which is beneficial for enhancing staff awareness and management from quintessence and outward manifestation of Japanese culture. By doing so,it will promote the cultural construction of Chinese agricultural enterprises and lift the core-competitiveness.

  17. Experimental Study on Self-assembly of KLD-12 Peptide Hydrogel and 3-D Culture of MSC Encapsulated within Hydrogel In Vitro

    Institute of Scientific and Technical Information of China (English)

    Jianhua SUN; Qixin ZHENG

    2009-01-01

    o-fiber hydrogel in vitro. MSCs in KLD-12 peptide hydrogel grew well and proliferated with the culture time. KLD-12 peptide hydrogel can serve as an excellent injectable material of biological scaffolds in tissue engineering of IVD.

  18. Preparation of cultured cells using high-pressure freezing and freeze substitution for subsequent 2D or 3D visualization in the transmission electron microscope.

    Science.gov (United States)

    Hawes, Philippa C

    2015-01-01

    Transmission electron microscopy (TEM) is an invaluable technique used for imaging the ultrastructure of samples and it is particularly useful when determining virus-host interactions at a cellular level. The environment inside a TEM is not favorable for biological material (high vacuum and high energy electrons). Also biological samples have little or no intrinsic electron contrast, and rarely do they naturally exist in very thin sheets, as is required for optimum resolution in the TEM. To prepare these samples for imaging in the TEM therefore requires extensive processing which can alter the ultrastructure of the material. Here we describe a method which aims to minimize preparation artifacts by freezing the samples at high pressure to instantaneously preserve ultrastructural detail, then rapidly substituting the ice and infiltrating with resin to provide a firm matrix which can be cut into thin sections for imaging. Thicker sections of this material can also be imaged and reconstructed into 3D volumes using electron tomography.

  19. Development, Characterization and Cell Cultural Response of 3D Biocompatible Micro-Patterned Poly-ε-Caprolactone Scaffolds Designed and Fabricated Integrating Lithography and Micromolding Fabrication Techniques

    KAUST Repository

    Limongi, Tania

    2014-12-12

    Scaffold design and fabrication are very important subjects for biomaterial, tissue engineering and regenerative medicine research playing a unique role in tissue regeneration and repair. Among synthetic biomaterials Poly-ε- Caprolactone (PCL) is very attractive bioresorbable polyester due to its high permeability, biodegradability and capacity to be blended with other biopolymers. Thanks to its ability to naturally degrade in tissues, PCL has a great potential as a new material for implantable biomedical micro devices. This work focuses on the establishment of a micro fabrication process, by integrating lithography and micromolding fabrication techniques, for the realization of 3D microstructure PCL devices. Scaffold surface exhibits a combination in the patterned length scale; cylindrical pillars of 10 μm height and 10 μm diameter are arranged in a hexagonal lattice with periodicity of 30 μm and their sidewalls are nano-sculptured, with a regular pattern of grooves leading to a spatial modulation in the z direction. In order to demonstrate that these biocompatible pillared PCL substrates are suitable for a proper cell growth, NIH/3T3 mouse embryonic fibroblasts were seeded on them and cells key adhesion parameters were evaluated. Scanning Electron Microscopy and immunofluorescence analysis were carried out to check cell survival, proliferation and adhesion; cells growing on the PCL substrates appeared healthy and formed a well-developed network in close contact with the micro and nano features of the pillared surface. Those 3D scaffolds could be a promising solution for a wide range of applications within tissue engineering and regenerative medicine applications.

  20. Direct measurement of dust attenuation in z~1.5 star-forming galaxies from 3D-HST: Implications for dust geometry and star formation rates

    CERN Document Server

    Price, Sedona H; Brammer, Gabriel B; Conroy, Charlie; Schreiber, Natascha M Forster; Franx, Marijn; Fumagalli, Mattia; Lundgren, Britt; Momcheva, Ivelina; Nelson, Erica J; Rix, Hans-Walter; Skelton, Rosalind E; van Dokkum, Pieter G; Whitaker, Katherine E; Wuyts, Stijn

    2013-01-01

    The nature of dust in distant galaxies is not well understood, and until recently few direct dust measurements have been possible. We investigate dust in distant star-forming galaxies using near-infrared grism spectra of the 3D-HST survey combined with archival multi-wavelength photometry. These data allow us to make a direct comparison between dust towards star-forming regions (measured using Balmer decrements) and the integrated dust properties (derived by comparing spectral energy distributions [SEDs] with stellar population and dust models) for a statistically significant sample of distant galaxies. We select a sample of 163 galaxies between 1.36 $\\le$ z $\\le$ 1.5 with H$\\alpha$ SNR $\\ge$ 5 and measure Balmer decrements from stacked spectra. First, we stack spectra in bins of integrated stellar dust attenuation, and find that there is extra dust extinction towards star-forming regions ($A_{V,HII}$ is 1.81 times the integrated $A_{V,star}$), though slightly lower than found for low-redshift starburst galax...

  1. Satellite-based 3D structure of cloud and aerosols over the Indian Monsoon region: implications for aerosol-cloud interaction

    Science.gov (United States)

    Dey, Sagnik; Sengupta, Kamalaika; Basil, George; Das, Sushant; Nidhi, Nidhi; Dash, S. K.; Sarkar, Arjya; Srivastava, Parul; Singh, Ajit; Agarwal, P.

    2012-11-01

    Accurate knowledge of vertical distributions of aerosol and cloud fields and their space-time variations are required to reduce the uncertainty in estimated climate forcing. Here, multi-sensor (both passive and active) data were used to construct the climatology of 3-D cloud and aerosol fields over the Indian monsoon region. Multilayer clouds are found to persist throughout the year, among which cumulus and stratocumulus dominate the low clouds and cirrus dominates the high clouds. A combination of passive stereo-technique (MISR) and radiometric technique (ISCPP) captures the multilayer cloud structure as revealed by active sensor CALIOP. Coexistence of low clouds throughout the year with high aerosol concentration beneath and above leads to a transition from increasing to decreasing cloud fraction with an increase in aerosol optical depth. Such transition is rapid in the monsoon season due to convergence of low clouds to form high clouds facilitated by high aerosol loading. Further, the regional climate model RegCM 4.1 has been used to examine aerosol-cloud interaction. The aerosol-induced changes of low cloud amount are under-estimated by the model. The observation-based seasonal climatology of aerosol and cloud fields presented here may help in improving the model simulations of cloud variability and associated rainfall.

  2. 3D-kompositointi

    OpenAIRE

    Piirainen, Jere

    2015-01-01

    Opinnäytetyössä käydään läpi yleisimpiä 3D-kompositointiin liittyviä tekniikoita sekä kompositointiin käytettyjä ohjelmia ja liitännäisiä. Työssä esitellään myös kompositoinnin juuret 1800-luvun lopulta aina nykyaikaiseen digitaaliseen kompositointiin asti. Kompositointi on yksinkertaisimmillaan usean kuvan liittämistä saumattomasti yhdeksi uskottavaksi kokonaisuudeksi. Vaikka prosessi vaatii visuaalista silmää, vaatii se myös paljon teknistä osaamista. Tämän lisäksi perusymmärrys kamera...

  3. Shaping 3-D boxes

    DEFF Research Database (Denmark)

    Stenholt, Rasmus; Madsen, Claus B.

    2011-01-01

    Enabling users to shape 3-D boxes in immersive virtual environments is a non-trivial problem. In this paper, a new family of techniques for creating rectangular boxes of arbitrary position, orientation, and size is presented and evaluated. These new techniques are based solely on position data......, making them different from typical, existing box shaping techniques. The basis of the proposed techniques is a new algorithm for constructing a full box from just three of its corners. The evaluation of the new techniques compares their precision and completion times in a 9 degree-of-freedom (Do......F) docking experiment against an existing technique, which requires the user to perform the rotation and scaling of the box explicitly. The precision of the users' box construction is evaluated by a novel error metric measuring the difference between two boxes. The results of the experiment strongly indicate...

  4. 3D Application Study

    Science.gov (United States)

    1989-11-01

    accuracy or confusion as to the actual scale of objects in the scene. Man-made objects representing fixed cultural features are subject to many of...4.2.1.8 Pepper’s Ghost This is a commercially available embodiment of holographic technology that is used at The Haunted Mansion in Disneyland . The... cultural features were not available to the demonstration implementation team, it was necessary to create entities that appear on the landscape. As

  5. A method for the design of 3D scaffolds for high-density cell attachment and determination of optimum perfusion culture conditions.

    Science.gov (United States)

    Provin, Christophe; Takano, Kiyoshi; Sakai, Yasuyuki; Fujii, Teruo; Shirakashi, Ryo

    2008-01-01

    The application of in vitro cultured cells in tissue engineering or drug screening, aimed at complex soft tissues such as liver, requires in vivo physiological function of the cultured cells. For this purpose, the scaffold in which cells are cultured should provide a microenvironment similar to an in vivo one with a three-dimensional extracellular matrix, a high supply capacity of O(2) and nutrients, and high cell density. In this paper, we propose a method to design (1) the geometry of the scaffold, with a surface/volume ratio optimized to allow high-density (5 x 10(7)cells/mL) cell culture and (2) culture conditions that will supply optimal quantities of oxygen and nutrients. CFD modeling of mass transport was used to determine the shear stress as well as O(2) and glucose metabolism in the scaffold (20 mm width-35 mm length) for various flow rates. Validation of the model was done through comparison with flow resistance and micro-PIV experiments. CFD analysis showed the maximum metabolic rate densities for this scaffold are 6.04 x 10(-3)mol/s/m(3) for O(2) at 0.71 mL/min and 1.91 x 10(-2)mol/s/m(3) for glucose at 0.35 mL/min.

  6. Epic Dimensions: a Comparative Analysis of 3d Acquisition Methods

    Science.gov (United States)

    Graham, C. A.; Akoglu, K. G.; Lassen, A. W.; Simon, S.

    2017-08-01

    When it comes to capturing the geometry of a cultural heritage artifact, there is certainly no dearth of possible acquisition techniques. As technology has rapidly developed, the availability of intuitive 3D generating tools has increased exponentially and made it possible even for non-specialists to create many models quickly. Though the by-products of these different acquisition methods may be incongruent in terms of quality, these discrepancies are not problematic, as there are many applications of 3D models, each with their own set of requirements. Comparisons of high-resolution 3D models of an iconic Babylonian tablet, captured via four different closerange technologies discussed in this paper assess which methods of 3D digitization best suit specific intended purposes related to research, conservation and education. Taking into consideration repeatability, time and resource implications, qualitative and quantitative potential and ease of use, this paper presents a study of the strengths and weakness of structured light scanning, triangulation laser scanning, photometric stereo and close-range photogrammetry, in the context of interactive investigation, conditions monitoring, engagement, and dissemination.

  7. Enzymatically degradable poly(ethylene glycol) hydrogels for the 3D culture and release of human embryonic stem cell derived pancreatic precursor cell aggregates.

    Science.gov (United States)

    Amer, Luke D; Holtzinger, Audrey; Keller, Gordon; Mahoney, Melissa J; Bryant, Stephanie J

    2015-08-01

    This study aimed to develop a three dimensional culture platform for aggregates of human embryonic stem cell (hESC)-derived pancreatic progenitors that enables long-term culture, maintains aggregate size and morphology, does not adversely affect differentiation and provides a means for aggregate recovery. A platform was developed with poly(ethylene glycol) hydrogels containing collagen type I, for cell-matrix interactions, and peptide crosslinkers, for facile recovery of aggregates. The platform was first demonstrated with RIN-m5F cells, showing encapsulation and subsequent release of single cells and aggregates without adversely affecting viability. Aggregates of hESC-derived pancreatic progenitors with an effective diameter of 82 (15)μm were either encapsulated in hydrogels or cultured in suspension for 28 days. At day 14, aggregate viability was maintained in the hydrogels, but significantly reduced (88%) in suspension culture. However by day 28, viability was reduced under both culture conditions. Aggregate size was maintained in the hydrogels, but in suspension was significantly higher (∼ 2-fold) by day 28. The ability to release aggregates followed by a second enzyme treatment to achieve single cells enabled assessment by flow cytometry. Prior to encapsulation, there were 39% Pdx1(+)/Nkx6.1(+) cells, key endocrine markers required for β-cell maturation. The fraction of doubly positive cells was not affected in hydrogels but was slightly and significantly lower in suspension culture by 28 days. In conclusion, we demonstrate that a MMP-sensitive PEG hydrogel containing collagen type I is a promising platform for hESC-derived pancreatic progenitors that maintains viable aggregates, aggregate size, and progenitor state and offers facile recovery of aggregates.

  8. 3D Printed Bionic Ears

    Science.gov (United States)

    Mannoor, Manu S.; Jiang, Ziwen; James, Teena; Kong, Yong Lin; Malatesta, Karen A.; Soboyejo, Winston O.; Verma, Naveen; Gracias, David H.; McAlpine, Michael C.

    2013-01-01

    The ability to three-dimensionally interweave biological tissue with functional electronics could enable the creation of bionic organs possessing enhanced functionalities over their human counterparts. Conventional electronic devices are inherently two-dimensional, preventing seamless multidimensional integration with synthetic biology, as the processes and materials are very different. Here, we present a novel strategy for overcoming these difficulties via additive manufacturing of biological cells with structural and nanoparticle derived electronic elements. As a proof of concept, we generated a bionic ear via 3D printing of a cell-seeded hydrogel matrix in the precise anatomic geometry of a human ear, along with an intertwined conducting polymer consisting of infused silver nanoparticles. This allowed for in vitro culturing of cartilage tissue around an inductive coil antenna in the ear, which subsequently enables readout of inductively-coupled signals from cochlea-shaped electrodes. The printed ear exhibits enhanced auditory sensing for radio frequency reception, and complementary left and right ears can listen to stereo audio music. Overall, our approach suggests a means to intricately merge biologic and nanoelectronic functionalities via 3D printing. PMID:23635097

  9. 3D printed bionic ears.

    Science.gov (United States)

    Mannoor, Manu S; Jiang, Ziwen; James, Teena; Kong, Yong Lin; Malatesta, Karen A; Soboyejo, Winston O; Verma, Naveen; Gracias, David H; McAlpine, Michael C

    2013-06-12

    The ability to three-dimensionally interweave biological tissue with functional electronics could enable the creation of bionic organs possessing enhanced functionalities over their human counterparts. Conventional electronic devices are inherently two-dimensional, preventing seamless multidimensional integration with synthetic biology, as the processes and materials are very different. Here, we present a novel strategy for overcoming these difficulties via additive manufacturing of biological cells with structural and nanoparticle derived electronic elements. As a proof of concept, we generated a bionic ear via 3D printing of a cell-seeded hydrogel matrix in the anatomic geometry of a human ear, along with an intertwined conducting polymer consisting of infused silver nanoparticles. This allowed for in vitro culturing of cartilage tissue around an inductive coil antenna in the ear, which subsequently enables readout of inductively-coupled signals from cochlea-shaped electrodes. The printed ear exhibits enhanced auditory sensing for radio frequency reception, and complementary left and right ears can listen to stereo audio music. Overall, our approach suggests a means to intricately merge biologic and nanoelectronic functionalities via 3D printing.

  10. The deep crustal structure of the mafic-ultramafic Seiland Igneous Province of Norway from 3-D gravity modelling and geological implications

    Science.gov (United States)

    Pastore, Zeudia; Fichler, Christine; McEnroe, Suzanne A.

    2016-12-01

    The Seiland Igneous Province (SIP) is the largest complex of mafic and ultramafic intrusions in northern Fennoscandia intruded at ca. 580-560 Ma. The depth extent and the deep structure of the SIP are mainly unknown apart from three profiles modelled by gravity and refraction seismic data. Utilizing 3-D gravity modelling, a complex model of the deep subsurface structure of the SIP has been developed. The structure is presented in a multiprofile model ranging from the surface to the Moho. The mafic/ultramafic rocks of the SIP are modelled with densities of 3100 and 3300 kg m-3, the surrounding rocks by densities of 2700 and 2900 kg m-3 for upper and lower crust, respectively. This density model explains the pronounced positive Bouguer gravity anomaly of up to 100 mGal above background. Its minimum volume is estimated from the subsurface model to 17 000 km3 and as such we revise downwards the earlier estimations of 25 000 km3. The new subsurface model suggests that most of the SIP has a thickness between 2 and 4 km. An area with roots in an annular pattern is found and two deep-reaching roots have been identified located below the islands of Seiland and Sørøy. The depth of these roots is estimated to approximatively 9 km. The SIP is presently interpreted to be in the Caledonian Kalak Nappe Complex and the roots depth constrains its minimum thickness which is larger than earlier estimated. Furthermore, the rather undisturbed shape of the annular root pattern indicates that the SIP has not been subjected to strong tectonic reworking during the Caledonian orogeny.

  11. Direct measurements of dust attenuation in z ∼ 1.5 star-forming galaxies from 3D-HST: Implications for dust geometry and star formation rates

    Energy Technology Data Exchange (ETDEWEB)

    Price, Sedona H.; Kriek, Mariska [Astronomy Department, University of California, Berkeley, CA 94720 (United States); Brammer, Gabriel B. [Space Telescope Science Institute, 3700 San Martin Drive, Baltimore, MD 21218 (United States); Conroy, Charlie [Department of Astronomy and Astrophysics, University of California, Santa Cruz, CA 95064 (United States); Schreiber, Natascha M. Förster; Wuyts, Stijn [Max-Planck-Institut für extraterrestrische Physik, Giessenbachstrasse, D-85748 Garching (Germany); Franx, Marijn; Fumagalli, Mattia [Leiden Observatory, Leiden University, P.O. Box 9513, 2300 RA Leiden (Netherlands); Lundgren, Britt [Department of Astronomy, University of Wisconsin, 475 N Charter Street, Madison, WI 53706 (United States); Momcheva, Ivelina; Nelson, Erica J.; Van Dokkum, Pieter G. [Department of Astronomy, Yale University, New Haven, CT 06511 (United States); Skelton, Rosalind E. [South African Astronomical Observatory, P.O. Box 9, Observatory 7935 (South Africa); Whitaker, Katherine E., E-mail: sedona@berkeley.edu [Astrophysics Science Division, Goddard Space Flight Center, Code 665, Greenbelt, MD 20771 (United States)

    2014-06-10

    The nature of dust in distant galaxies is not well understood, and until recently few direct dust measurements have been possible. We investigate dust in distant star-forming galaxies using near-infrared grism spectra of the 3D-HST survey combined with archival multi-wavelength photometry. These data allow us to make a direct comparison between dust around star-forming regions (A {sub V,} {sub H} {sub II}) and the integrated dust content (A {sub V,} {sub star}). We select a sample of 163 galaxies between 1.36 ≤ z ≤ 1.5 with Hα signal-to-noise ratio ≥5 and measure Balmer decrements from stacked spectra to calculate A {sub V,} {sub H} {sub II}. First, we stack spectra in bins of A {sub V,} {sub star}, and find that A {sub V,} {sub H} {sub II} = 1.86 A {sub V,} {sub star}, with a significance of σ = 1.7. Our result is consistent with the two-component dust model, in which galaxies contain both diffuse and stellar birth cloud dust. Next, we stack spectra in bins of specific star formation rate (log SSFR), star formation rate (log SFR), and stellar mass (log M {sub *}). We find that on average A {sub V,} {sub H} {sub II} increases with SFR and mass, but decreases with increasing SSFR. Interestingly, the data hint that the amount of extra attenuation decreases with increasing SSFR. This trend is expected from the two-component model, as the extra attenuation will increase once older stars outside the star-forming regions become more dominant in the galaxy spectrum. Finally, using Balmer decrements we derive dust-corrected Hα SFRs, and find that stellar population modeling produces incorrect SFRs if rapidly declining star formation histories are included in the explored parameter space.

  12. Direct Measurement of Dust Attenuation in z approx. 1.5 Star-Forming Galaxies from 3D-HST: Implications for Dust Geometry and Star Formation Rates

    Science.gov (United States)

    Price, Sedona H.; Kriek, Mariska; Brammer, Gabriel B; Conroy, Charlie; Schreiber, Natascha M. Foerster; Franx, Marijn; Fumagalli, Mattia; Lundren, Britt; Momcheva, Ivelina; Nelson, Erica J.; Rix, Hans-Walter; Skelton, Rosalind E.; VanDokkum, Pieter G.; Tease, Katherine Whitaker; Wuyts, Stijn

    2013-01-01

    The nature of dust in distant galaxies is not well understood, and until recently few direct dust measurements have been possible. We investigate dust in distant star-forming galaxies using near-infrared grism spectra of the 3D-HST survey combined with archival multi-wavelength photometry. These data allow us to make a direct comparison between dust towards star-forming regions (measured using Balmer decrements) and the integrated dust properties (derived by comparing spectral energy distributions [SEDs] with stellar population and dust models) for a statistically significant sample of distant galaxies. We select a sample of 163 galaxies between 1.36 or = 5 and measure Balmer decrements from stacked spectra. First, we stack spectra in bins of integrated stellar dust attenuation, and find that there is extra dust extinction towards star-forming regions (AV,HII is 1.81 times the integrated AV, star), though slightly lower than found for low-redshift starburst galaxies. Next, we stack spectra in bins of specific star formation rate (log sSFR), star formation rate (log SFR), and stellar mass (logM*). We find that on average AV,HII increases with SFR and mass, but decreases with increasing sSFR. The amount of extra extinction also decreases with increasing sSFR and decreasing stellar mass. Our results are consistent with the two-phase dust model - in which galaxies contain both a diffuse and a stellar birth cloud dust component - as the extra extinction will increase once older stars outside the star-forming regions become more dominant. Finally, using our Balmer decrements we derive dust-corrected H(alpha) SFRs, and find evidence that SED fitting produces incorrect SFRs if very rapidly declining SFHs are included in the explored parameter space. Subject headings: dust, extinction- galaxies: evolution- galaxies: high-redshift

  13. 3-D GPS velocity field and its implications on the present-day post-orogenic deformation of the Western Alps and Pyrenees

    Science.gov (United States)

    Ninh Nguyen, Hai; Vernant, Philippe; Mazzotti, Stephane; Khazaradze, Giorgi; Asensio, Eva

    2016-09-01

    We present a new 3-D GPS velocity solution for 182 sites for the region encompassing the Western Alps, Pyrenees, and southern France. The velocity field is based on a Precise Point Positioning (PPP) solution, to which we apply a common-mode filter, defined by the 26 longest time series, in order to correct for network-wide biases (reference frame, unmodeled large-scale processes, etc.). We show that processing parameters, such as troposphere delay modeling, can lead to systematic velocity variations of 0.1-0.5 mm yr-1 affecting both accuracy and precision, especially for short (< 5 years) time series. A velocity convergence analysis shows that minimum time-series lengths of ˜ 3 and ˜ 5.5 years are required to reach a velocity stability of 0.5 mm yr-1 in the horizontal and vertical components, respectively. On average, horizontal residual velocities show a stability of ˜ 0.2 mm yr-1 in the Western Alps, Pyrenees, and southern France. The only significant horizontal strain rate signal is in the western Pyrenees with up to 4 × 10-9 yr-1 NNE-SSW extension, whereas no significant strain rates are detected in the Western Alps (< 1 × 10-9 yr-1). In contrast, we identify significant uplift rates up to 2 mm yr-1 in the Western Alps but not in the Pyrenees (0.1 ± 0.2 mm yr-1). A correlation between site elevations and fast uplift rates in the northern part of the Western Alps, in the region of the Würmian ice cap, suggests that part of this uplift is induced by postglacial rebound. The very slow uplift rates in the southern Western Alps and in the Pyrenees could be accounted for by erosion-induced rebound.

  14. Analysis of Gene Expression Signatures for Osteogenic 3D Perfusion-Bioreactor Cell Cultures Based on a Multifactorial DoE Approach

    Directory of Open Access Journals (Sweden)

    Ioannis Papantoniou

    2014-08-01

    Full Text Available The use of multifactorial design of experiments (DoE in tissue engineering bioprocess development will contribute to the robust manufacturing of tissue engineered constructs by linking their quality characteristics to bioprocess operating parameters. In this work, perfusion bioreactors were used for the in vitro culture and osteogenic differentiation of human periosteum-derived cells (hPDCs seeded on three-dimensional titanium (Ti alloy scaffolds. A CaP-supplemented medium was used to induce differentiation of the cultured hPDCs. A two-level, three-factor fractional factorial design was employed to evaluate a range of bioreactor operating conditions by changing the levels of the following parameters: flow rate (0.5–2 mL/min, cell culture duration (7–21 days and cell seeding density (1.5 × 103–3 × 103 cells/cm2. This approach allowed for evaluating the individual impact of the aforementioned process parameters upon a range of genes that are related to the osteogenic lineage, such as collagen type I, alkaline phosphatase, osterix, osteopontin and osteocalcin. Furthermore, by overlaying gene-specific response surfaces, an integrated operating process space was highlighted within which predetermined values of the six genes of interest (i.e., gene signature could be minimally met over the course of the bioreactor culture time.

  15. Conducting Polymer 3D Microelectrodes

    Directory of Open Access Journals (Sweden)

    Jenny Emnéus

    2010-12-01

    Full Text Available Conducting polymer 3D microelectrodes have been fabricated for possible future neurological applications. A combination of micro-fabrication techniques and chemical polymerization methods has been used to create pillar electrodes in polyaniline and polypyrrole. The thin polymer films obtained showed uniformity and good adhesion to both horizontal and vertical surfaces. Electrodes in combination with metal/conducting polymer materials have been characterized by cyclic voltammetry and the presence of the conducting polymer film has shown to increase the electrochemical activity when compared with electrodes coated with only metal. An electrochemical characterization of gold/polypyrrole electrodes showed exceptional electrochemical behavior and activity. PC12 cells were finally cultured on the investigated materials as a preliminary biocompatibility assessment. These results show that the described electrodes are possibly suitable for future in-vitro neurological measurements.

  16. Intraoral 3D scanner

    Science.gov (United States)

    Kühmstedt, Peter; Bräuer-Burchardt, Christian; Munkelt, Christoph; Heinze, Matthias; Palme, Martin; Schmidt, Ingo; Hintersehr, Josef; Notni, Gunther

    2007-09-01

    Here a new set-up of a 3D-scanning system for CAD/CAM in dental industry is proposed. The system is designed for direct scanning of the dental preparations within the mouth. The measuring process is based on phase correlation technique in combination with fast fringe projection in a stereo arrangement. The novelty in the approach is characterized by the following features: A phase correlation between the phase values of the images of two cameras is used for the co-ordinate calculation. This works contrary to the usage of only phase values (phasogrammetry) or classical triangulation (phase values and camera image co-ordinate values) for the determination of the co-ordinates. The main advantage of the method is that the absolute value of the phase at each point does not directly determine the coordinate. Thus errors in the determination of the co-ordinates are prevented. Furthermore, using the epipolar geometry of the stereo-like arrangement the phase unwrapping problem of fringe analysis can be solved. The endoscope like measurement system contains one projection and two camera channels for illumination and observation of the object, respectively. The new system has a measurement field of nearly 25mm × 15mm. The user can measure two or three teeth at one time. So the system can by used for scanning of single tooth up to bridges preparations. In the paper the first realization of the intraoral scanner is described.

  17. Martian terrain - 3D

    Science.gov (United States)

    1997-01-01

    This area of terrain near the Sagan Memorial Station was taken on Sol 3 by the Imager for Mars Pathfinder (IMP). 3D glasses are necessary to identify surface detail.The IMP is a stereo imaging system with color capability provided by 24 selectable filters -- twelve filters per 'eye.' It stands 1.8 meters above the Martian surface, and has a resolution of two millimeters at a range of two meters.Mars Pathfinder is the second in NASA's Discovery program of low-cost spacecraft with highly focused science goals. The Jet Propulsion Laboratory, Pasadena, CA, developed and manages the Mars Pathfinder mission for NASA's Office of Space Science, Washington, D.C. JPL is an operating division of the California Institute of Technology (Caltech). The Imager for Mars Pathfinder (IMP) was developed by the University of Arizona Lunar and Planetary Laboratory under contract to JPL. Peter Smith is the Principal Investigator.Click below to see the left and right views individually. [figure removed for brevity, see original site] Left [figure removed for brevity, see original site] Right

  18. 3D Printing and 3D Bioprinting in Pediatrics.

    Science.gov (United States)

    Vijayavenkataraman, Sanjairaj; Fuh, Jerry Y H; Lu, Wen Feng

    2017-07-13

    Additive manufacturing, commonly referred to as 3D printing, is a technology that builds three-dimensional structures and components layer by layer. Bioprinting is the use of 3D printing technology to fabricate tissue constructs for regenerative medicine from cell-laden bio-inks. 3D printing and bioprinting have huge potential in revolutionizing the field of tissue engineering and regenerative medicine. This paper reviews the application of 3D printing and bioprinting in the field of pediatrics.

  19. Arginine-glycine-aspartic acid-polyethylene glycol-polyamidoamine dendrimer conjugate improves liver-cell aggregation and function in 3-D spheroid culture.

    Science.gov (United States)

    Chen, Zhanfei; Lian, Fen; Wang, Xiaoqian; Chen, Yanling; Tang, Nanhong

    The polyamidoamine (PAMAM) dendrimer, a type of macromolecule material, has been used in spheroidal cell culture and drug delivery in recent years. However, PAMAM is not involved in the study of hepatic cell-spheroid culture or its biological activity, particularly in detoxification function. Here, we constructed a PAMAM-dendrimer conjugate decorated by an integrin ligand: arginine-glycine-aspartic acid (RGD) peptide. Our studies demonstrate that RGD-polyethylene glycol (PEG)-PAMAM conjugates can promote singly floating hepatic cells to aggregate together in a sphere-like growth with a weak reactive oxygen species. Moreover, RGD-PEG-PAMAM conjugates can activate the AKT-MAPK pathway in hepatic cells to promote cell proliferation and improve basic function and ammonia metabolism. Together, our data support the hepatocyte sphere treated by RGD-PEG-PAMAM conjugates as a potential source of hepatic cells for a biological artificial liver system.

  20. Modular, pumpless body-on-a-chip platform for the co-culture of GI tract epithelium and 3D primary liver tissue.

    Science.gov (United States)

    Esch, Mandy B; Ueno, Hidetaka; Applegate, Dawn R; Shuler, Michael L

    2016-07-05

    We have developed an expandable modular body-on-a-chip system that allows for a plug-and-play approach with several in vitro tissues. The design consists of single-organ chips that are combined with each other to yield a multi-organ body-on-a-chip system. Fluidic flow through the organ chips is driven via gravity and controlled passively via hydraulic resistances of the microfluidic channel network. Such pumpless body-on-a-chip devices are inexpensive and easy to use. We tested the device by culturing GI tract tissue and liver tissue within the device. Integrated Ag/AgCl electrodes were used to measure the resistance across the GI tract cell layer. The transepithelial resistance (TEER) reached values between 250 to 650 Ω cm(2) throughout the 14 day co-culture period. These data indicate that the GI tract cells retained their viability and the GI tract layer as a whole retained its barrier function. Throughout the 14 day co-culture period we measured low amounts of aspartate aminotransferase (AST, ∼10-17.5 U L(-1)), indicating low rates of liver cell death. Metabolic rates of hepatocytes were comparable to those of hepatocytes in single-organ fluidic cell culture systems (albumin production ranged between 3-6 μg per day per million hepatocytes and urea production ranged between 150-200 μg per day per million hepatocytes). Induced CYP activities were higher than previously measured with microfluidic liver only systems.

  1. 3D printing for dummies

    CERN Document Server

    Hausman, Kalani Kirk

    2014-01-01

    Get started printing out 3D objects quickly and inexpensively! 3D printing is no longer just a figment of your imagination. This remarkable technology is coming to the masses with the growing availability of 3D printers. 3D printers create 3-dimensional layered models and they allow users to create prototypes that use multiple materials and colors.  This friendly-but-straightforward guide examines each type of 3D printing technology available today and gives artists, entrepreneurs, engineers, and hobbyists insight into the amazing things 3D printing has to offer. You'll discover methods for

  2. The effect of light-emitting diode irradiation at different wavelengths on calcification of osteoblast-like cells in 3D culture.

    Science.gov (United States)

    Chintavalakorn, Rochaya; Tanglitanont, Tatsanee; Khantachawana, Anak; Viravaidya-Pasuwat, Kwanchanok; Santiwong, Peerapong

    2015-08-01

    This study aimed to investigate the effect of four different light-emitting diode (LED) wavelengths on calcification and proliferation of osteoblast-like cells in vitro. MC3T3-E1 cells were seeded within three-dimensional collagen scaffolds and irradiated daily by LED light with peak emission wavelengths of 630-, 680-, 760- and 830-nm at constant fluency of 3.1 J/cm(2) (irradiance intensity 2 mW/cm(2)). Cultures were measured for calcium content at day 0, 7, 14, 21, 28, 35 and 42. The significant enhancement in calcium content was observed at the early stage of culture (days 7 and 14) (plight irradiation on osteoblastic cell calcification. Only 680-nm irradiated samples revealed a significant enhancement of calcium content until the late stages of culture (from days 21 to 42) (p<;0.001). The cyclin D mRNA expression that was investigated 3 hours after stimulation at day3 also show that the 680-nm LED irradiation can enhance cyclin D expression more than others. For enhancing bone mineralization, LED irradiation at the 680-nm is more effective than those at 630-, 760- and 830-nm. Further studies should be investigated in order to obtain the most effective parameters of LLLI on bone regeneration in clinical setting.

  3. Arginine–glycine–aspartic acid–polyethylene glycol–polyamidoamine dendrimer conjugate improves liver-cell aggregation and function in 3-D spheroid culture

    Directory of Open Access Journals (Sweden)

    Chen Z

    2016-08-01

    Full Text Available Zhanfei Chen,1,* Fen Lian,1,* Xiaoqian Wang,1 Yanling Chen,1,2 Nanhong Tang1,2 1Fujian Institute of Hepatobiliary Surgery, Fujian Medical University Union Hospital, 2Key Laboratory of Ministry of Education for Gastrointestinal Cancer, Research Center for Molecular Medicine, Fujian Medical University, Fuzhou, People’s Republic of China *These authors contributed equally to this work Abstract: The polyamidoamine (PAMAM dendrimer, a type of macromolecule material, has been used in spheroidal cell culture and drug delivery in recent years. However, PAMAM is not involved in the study of hepatic cell-spheroid culture or its biological activity, particularly in detoxification function. Here, we constructed a PAMAM-dendrimer conjugate decorated by an integrin ligand: arginine–glycine–aspartic acid (RGD peptide. Our studies demonstrate that RGD–polyethylene glycol (PEG–PAMAM conjugates can promote singly floating hepatic cells to aggregate together in a sphere-like growth with a weak reactive oxygen species. Moreover, RGD-PEG-PAMAM conjugates can activate the AKT–MAPK pathway in hepatic cells to promote cell proliferation and improve basic function and ammonia metabolism. Together, our data support the hepatocyte sphere treated by RGD-PEG-PAMAM conjugates as a potential source of hepatic cells for a biological artificial liver system. Keywords: dendrimer, arginine–glycine–aspartic acid (RGD, liver cell, spheroid culture, ammonia metabolism

  4. 模拟微重力下三维培养对人肝干细胞增殖和分化的影响%Effects of 3-D simulated microgravity culture on human hepatic stem cell proliferation and differentiation

    Institute of Scientific and Technical Information of China (English)

    姚新宇; 曹阳; 杜兴冉; 章莉莉

    2011-01-01

    目的:探讨微重力环境对肝干细胞增殖及分化的影响。方法:从人肝组织中分离、培养肝干细胞,进行肝干细胞鉴定后,分别进行平面培养、微载体培养、模拟微重力下三维培养。利用倒置相差显微镜、扫描电镜观察不同培养条件下细胞形态学变化;检测上清液中葡萄糖消耗量和乳酸脱氢酶含量以比较细胞增殖和活性情况;荧光定量PCR法检测细胞ALB、EpCAM、CK19、AFP、HNF6、CYP3A4、CYP3A7 mRNA表达变化。结果:模拟微重力环境下,肝干细胞以微载体球面为基底呈三维立体结构生长,形成大小不等细胞团;扫描电镜下见细胞团表面有丰富的细胞外基质及微绒毛;从9 d开始葡萄糖消耗量明显增加,至17 d达高峰,而LDH含量始终保持低水平;培养至21 d细胞仍高表达EpCAM、ALB、CK19、HNF6、CYP3A7,不表达AFP、CYP3A4,符合肝干细胞表型特征,与微载体培养结果相比有统计学差异(P<0.05)。结论:模拟微重力环境能促进肝干细胞呈三维立体结构生长,有利于细胞快速增殖并维持细胞活性和表型。%Objective:To investigate the effects of 3-D simulated microgravity culture on human hepatic stem cell proliferation and differentiation. Methods:The human hepatic stem cells,which were isolated from human fetal livers,were seeded in simulated microgravity 3-D Rotary Cell Culture System,conventional static culture and microcarrier culture system respectively. Cell morphology and growth pattern were observed under inverted phase contrast microscope and scanning electron microscope (SEM). Cellular proliferative activity and viability were assessed by monitoring the glucose consumption and Iactate dehydrogenase activity in culture supernatants respectively. The specific transcripts of hepatic progenitors (ALB,EpCAM,CK19,AFP,HNF6,CYP3A4,CYP3A7) in different culture systems were detected by quantitative RT-PCR. Results

  5. 3D velocity structure of the outer forearc of the Colombia-Ecuador subduction zone; implications for the 1958 megathrust earthquake rupture zone

    Science.gov (United States)

    Galve, A.; Charvis, P.; Garcia Cano, L.; Marcaillou, B.

    2013-12-01

    In 2005, we conducted an onshore-offshore 3D refraction and wide-angle reflection seismic experiment over the rupture zone of the 1958 subduction earthquake that occurred near the border between Colombia and Ecuador. This earthquake was part of a sequence of 3 large ruptures (1942, Mw=7.8; 1958, Mw=7.7; 1979, Mw=8.2), which successively broke from south to north the segments of the megathrust that had been ruptured in 1906 by a single, very large magnitude (8.8) earthquake. Using first arrival traveltime inversion, we constructed a well-defined Vp velocity model of the plate boundary and of the upper and lower plates, down to 25 km depth. The model reveals a 5-km thick, low velocity zone in the upper plate, located immediately above the interplate contact. Because similar low-velocity zones are commonly observed along margins made of oceanic or island-arc accreted terranes, we suggest that the low-velocity zone might result from the alteration and hydration of mafic and ultramafic rocks in the upper plate basement, rather than from hydrofracturing alone. Sediments underplated beneath the inner wedge might contribute to the low-velocity zone but it is unlikely that they are several kilometers thick. Nevertheless, fluids expelled by the compaction and dehydration of those underplated sediments possibly favor the alteration of the overlying rocks. The low-velocity zone is spatially coincident with the 1958 rupture area. Near the toe of the margin, the model shows a low velocity gradient in the outer wedge that we interpret as a zone of highly faulted and fractured rocks or of poorly consolidated sediments. This low velocity/low gradient region forms the oceanward limit of the rupture zones of both the 1958 and the 1979 earthquakes. We suggest that the two earthquake ruptures were arrested by the low velocity zone because its low rigidity contributed to dissipate most of the seismic energy and of the coseismic strain/stress. This might be the reason why the 1958

  6. From "sapienza" to "sapienza, State Archives in Rome". a Looping Effect Bringing Back to the Original Source Comunication and Culture by Innovative and Low Cost 3d Surveying, Imaging Systems and GIS Applications

    Science.gov (United States)

    Paolini, P.; Forti, G.; Catalani, G.; Lucchetti, S.; Menghini, A.; Mirandola, A.; Pistacchio, S.; Porzia, U.; Roberti, M.

    2016-04-01

    High Quality survey models, realized by multiple Low Cost methods and technologies, as a container to sharing Cultural and Archival Heritage, this is the aim guiding our research, here described in its primary applications. The SAPIENZA building, a XVI century masterpiece that represented the first unified headquarters of University in Rome, plays since year 1936, when the University moved to its newly edified campus, the role of the main venue for the State Archives. By the collaboration of a group of students of the Architecture Faculty, some integrated survey methods were applied on the monument with success. The beginning was the topographic survey, creating a reference on ground and along the monument for the upcoming applications, a GNNS RTK survey followed georeferencing points on the internal courtyard. Dense stereo matching photogrammetry is nowadays an accepted method for generating 3D survey models, accurate and scalable; it often substitutes 3D laser scanning for its low cost, so that it became our choice. Some 360° shots were planned for creating panoramic views of the double portico from the courtyard, plus additional single shots of some lateral spans and of pillars facing the court, as a single operation with a double finality: to create linked panotours with hotspots to web-linked databases, and 3D textured and georeferenced surface models, allowing to study the harmonic proportions of the classical architectural order. The use of free web Gis platforms, to load the work in Google Earth and the realization of low cost 3D prototypes of some representative parts, has been even performed.

  7. FROM ”SAPIENZA” TO “SAPIENZA, STATE ARCHIVES IN ROME”. A LOOPING EFFECT BRINGING BACK TO THE ORIGINAL SOURCE COMUNICATION AND CULTURE BY INNOVATIVE AND LOW COST 3D SURVEYING, IMAGING SYSTEMS AND GIS APPLICATIONS

    Directory of Open Access Journals (Sweden)

    P. Paolini

    2016-04-01

    Full Text Available High Quality survey models, realized by multiple Low Cost methods and technologies, as a container to sharing Cultural and Archival Heritage, this is the aim guiding our research, here described in its primary applications. The SAPIENZA building, a XVI century masterpiece that represented the first unified headquarters of University in Rome, plays since year 1936, when the University moved to its newly edified campus, the role of the main venue for the State Archives. By the collaboration of a group of students of the Architecture Faculty, some integrated survey methods were applied on the monument with success. The beginning was the topographic survey, creating a reference on ground and along the monument for the upcoming applications, a GNNS RTK survey followed georeferencing points on the internal courtyard. Dense stereo matching photogrammetry is nowadays an accepted method for generating 3D survey models, accurate and scalable; it often substitutes 3D laser scanning for its low cost, so that it became our choice. Some 360° shots were planned for creating panoramic views of the double portico from the courtyard, plus additional single shots of some lateral spans and of pillars facing the court, as a single operation with a double finality: to create linked panotours with hotspots to web-linked databases, and 3D textured and georeferenced surface models, allowing to study the harmonic proportions of the classical architectural order. The use of free web Gis platforms, to load the work in Google Earth and the realization of low cost 3D prototypes of some representative parts, has been even performed.

  8. 3D VISUALIZATION FOR VIRTUAL MUSEUM DEVELOPMENT

    Directory of Open Access Journals (Sweden)

    M. Skamantzari

    2016-06-01

    Full Text Available The interest in the development of virtual museums is nowadays rising rapidly. During the last decades there have been numerous efforts concerning the 3D digitization of cultural heritage and the development of virtual museums, digital libraries and serious games. The realistic result has always been the main concern and a real challenge when it comes to 3D modelling of monuments, artifacts and especially sculptures. This paper implements, investigates and evaluates the results of the photogrammetric methods and 3D surveys that were used for the development of a virtual museum. Moreover, the decisions, the actions, the methodology and the main elements that this kind of application should include and take into consideration are described and analysed. It is believed that the outcomes of this application will be useful to researchers who are planning to develop and further improve the attempts made on virtual museums and mass production of 3D models.

  9. Networked 3D Virtual Museum System

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    Virtual heritage has become increasingly important in the conservation, preservation, and interpretation of our cultural and natural history. Moreover, rapid advances in digital technologies in recent years offer virtual heritage new direction. This paper introduces our approach toward a networked 3D virtual museum system, especially, how to model, manage, present virtual heritages and furthermore how to use computer network for the share of virtual heritage in the networked virtual environment. This paper first addresses a 3D acquisition and processing technique for virtual heritage modeling and shows some illustrative examples. Then, this paper describes a management of virtual heritage assets that are composed by various rich media. This paper introduces our schemes to present the virtual heritages, which include 3D virtual heritage browser system, CAVE system, and immersive VR theater. Finally, this paper presents the new direction of networked 3D virtual museum of which main idea is remote guide of the virtual heritage using the mixed reality technique.

  10. Evaluating the social and cultural implications of the internet

    OpenAIRE

    Brey, Philip

    2006-01-01

    Since the Internet's breakthrough as a mass medium, it has become a topic of discussion because of its implications for society. At one extreme, one finds those who only see great benefits and consider the Internet a tool for freedom, commerce, connectivity, and other societal benefits. At the other extreme, one finds those who lament the harms and disadvantages of the Internet, and who consider it a grave danger to existing social structures and institutions, to culture, morality and human r...

  11. 3D game environments create professional 3D game worlds

    CERN Document Server

    Ahearn, Luke

    2008-01-01

    The ultimate resource to help you create triple-A quality art for a variety of game worlds; 3D Game Environments offers detailed tutorials on creating 3D models, applying 2D art to 3D models, and clear concise advice on issues of efficiency and optimization for a 3D game engine. Using Photoshop and 3ds Max as his primary tools, Luke Ahearn explains how to create realistic textures from photo source and uses a variety of techniques to portray dynamic and believable game worlds.From a modern city to a steamy jungle, learn about the planning and technological considerations for 3D modelin

  12. A microfluidic device for 2D to 3D and 3D to 3D cell navigation

    Science.gov (United States)

    Shamloo, Amir; Amirifar, Leyla

    2016-01-01

    Microfluidic devices have received wide attention and shown great potential in the field of tissue engineering and regenerative medicine. Investigating cell response to various stimulations is much more accurate and comprehensive with the aid of microfluidic devices. In this study, we introduced a microfluidic device by which the matrix density as a mechanical property and the concentration profile of a biochemical factor as a chemical property could be altered. Our microfluidic device has a cell tank and a cell culture chamber to mimic both 2D to 3D and 3D to 3D migration of three types of cells. Fluid shear stress is negligible on the cells and a stable concentration gradient can be obtained by diffusion. The device was designed by a numerical simulation so that the uniformity of the concentration gradients throughout the cell culture chamber was obtained. Adult neural cells were cultured within this device and they showed different branching and axonal navigation phenotypes within varying nerve growth factor (NGF) concentration profiles. Neural stem cells were also cultured within varying collagen matrix densities while exposed to NGF concentrations and they experienced 3D to 3D collective migration. By generating vascular endothelial growth factor concentration gradients, adult human dermal microvascular endothelial cells also migrated in a 2D to 3D manner and formed a stable lumen within a specific collagen matrix density. It was observed that a minimum absolute concentration and concentration gradient were required to stimulate migration of all types of the cells. This device has the advantage of changing multiple parameters simultaneously and is expected to have wide applicability in cell studies.

  13. Changes in gene expression, protein content and morphology of chondrocytes cultured on a 3D Random Positioning Machine and 2D rotating clinostat

    Science.gov (United States)

    Aleshcheva, Ganna; Hauslage, Jens; Hemmersbach, Ruth; Infanger, Manfred; Bauer, Johann; Grimm, Daniela; Sahana, Jayashree

    Chondrocytes are the only cell type found in human cartilage consisting of proteoglycans and type II collagen. Several studies on chondrocytes cultured either in Space or on a ground-based facility for simulation of microgravity revealed that these cells are very resistant to adverse effects and stress induced by altered gravity. Tissue engineering of chondrocytes is a new strategy for cartilage regeneration. Using a three-dimensional Random Positioning Machine and a 2D rotating clinostat, devices designed to simulate microgravity on Earth, we investigated the early effects of microgravity exposure on human chondrocytes of six different donors after 30 min, 2 h, 4 h, 16 h, and 24 h and compared the results with the corresponding static controls cultured under normal gravity conditions. As little as 30 min of exposure resulted in increased expression of several genes responsible for cell motility, structure and integrity (beta-actin); control of cell growth, cell proliferation, cell differentiation and apoptosis; and cytoskeletal components such as microtubules (beta-tubulin) and intermediate filaments (vimentin). After 4 hours disruptions in the vimentin network were detected. These changes were less dramatic after 16 hours, when human chondrocytes appeared to reorganize their cytoskeleton. However, the gene expression and protein content of TGF-β1 was enhanced for 24 h. Based on the results achieved, we suggest that chondrocytes exposed to simulated microgravity seem to change their extracellular matrix production behavior while they rearrange their cytoskeletal proteins prior to forming three-dimensional aggregates.

  14. 3D Printing an Octohedron

    OpenAIRE

    Aboufadel, Edward F.

    2014-01-01

    The purpose of this short paper is to describe a project to manufacture a regular octohedron on a 3D printer. We assume that the reader is familiar with the basics of 3D printing. In the project, we use fundamental ideas to calculate the vertices and faces of an octohedron. Then, we utilize the OPENSCAD program to create a virtual 3D model and an STereoLithography (.stl) file that can be used by a 3D printer.

  15. Salient Local 3D Features for 3D Shape Retrieval

    CERN Document Server

    Godil, Afzal

    2011-01-01

    In this paper we describe a new formulation for the 3D salient local features based on the voxel grid inspired by the Scale Invariant Feature Transform (SIFT). We use it to identify the salient keypoints (invariant points) on a 3D voxelized model and calculate invariant 3D local feature descriptors at these keypoints. We then use the bag of words approach on the 3D local features to represent the 3D models for shape retrieval. The advantages of the method are that it can be applied to rigid as well as to articulated and deformable 3D models. Finally, this approach is applied for 3D Shape Retrieval on the McGill articulated shape benchmark and then the retrieval results are presented and compared to other methods.

  16. Type conversion of secretomes in a 3D TAM2 and HCC cell co-culture system and functional importance of CXCL2 in HCC

    Science.gov (United States)

    Lu, Yu; Li, Shan; Ma, Liping; Li, Yan; Zhang, Xiaolian; Peng, Qiliu; Mo, Cuiju; Huang, Li; Qin, Xue; Liu, Yinkun

    2016-01-01

    Macrophages play important roles in the tumor microenvironment, driving cancer progression and metastasis, particularly in hepatocellular carcinoma (HCC). However, few studies have assessed the exact secretome composition in HCC. In the present study, the impact of different phenotype of macrophages on HCC cells was investigated. Alternatively activated macrophages (M2) were found to significantly increase the proliferation, migration, and invasion abilities of SMMC7721 cells (all P 1.3-fold) and 96 down-regulated (<0.7-fold). CXCL2 was confirmed to have higher expression in the co-culture system and HCC tissues, and was selected for further investigation. Functional effects data suggested that recombinant human CXCL2 significantly enhanced the migration, invasion ability of SMMC7721 cells, and weakened adhesion ability. While CXCL2 neutralization and CXCR2 blockage significantly inhibited the effects of CXCL2 on SMMC7721 cells, indicating that CXCL2 may play pivotal role in HCC metastasis. PMID:27117207

  17. Characterization of the in vitro propagation of epileptiform electrophysiological activity in organotypic hippocampal slice cultures coupled to 3D microelectrode arrays

    DEFF Research Database (Denmark)

    Pisciotta, Marzia; Morgavi, Giovanna; Jahnsen, Henrik

    2010-01-01

    Dynamic aspects of the propagation of epileptiform activity have so far received little attention. With the aim of providing new insights about the spatial features of the propagation of epileptic seizures in the nervous system, we studied in vitro the initiation and propagation of traveling epil......AnimalsAnimals, NewbornConvulsants/pharmacologyElectric Stimulation/methodsElectrophysiological Phenomena/drug effectsElectrophysiological Phenomena/physiology*Evoked Potentials/drug effectsEvoked Potentials/physiology*Hippocampus/anatomy & histologyHippocampus/drug effects......Hippocampus/physiology*Microelectrodes*Organ Culture TechniquesPicrotoxin/pharmacologyRatsRats, WistarReaction Time/drug effectsReaction Time/physiologyTime FactorsSubstancesConvulsantsPicrotoxin LinkOut - more resourcesFull Text SourcesElsevier ScienceEBSCOOhioLINK Electronic Journal CenterSwets Information ServicesMolecular Biology Databases...

  18. The organic osmolyte betaine induces keratin 2 expression in rat epidermal keratinocytes - A genome-wide study in UVB irradiated organotypic 3D cultures.

    Science.gov (United States)

    Rauhala, Leena; Hämäläinen, Lasse; Dunlop, Thomas W; Pehkonen, Petri; Bart, Geneviève; Kokkonen, Maarit; Tammi, Markku; Tammi, Raija; Pasonen-Seppänen, Sanna

    2015-12-25

    The moisturizing and potentially protective properties of the organic osmolyte betaine (trimethylglycine) have made it an attractive component for skin care products. Its wide use despite the lack of comprehensive studies addressing its specific effects in skin led us to characterize the molecular targets of betaine in keratinocytes and to explore, whether it modifies the effects of acute UVB exposure. Genome-wide expression analysis was performed on organotypic cultures of rat epidermal keratinocytes, treated either with betaine (10mM), UVB (30 mJ/cm(2)) or their combination. Results were verified with qRT-PCR, western blotting and immunohistochemistry. Additionally, cell proliferation and differentiation were analyzed. Among the 89 genes influenced by betaine, the differentiation marker keratin 2 showed the highest upregulation, which was also confirmed at protein level. Expression of Egr1, a transcription factor, and Purkinje cell protein 4, a regulator of Ca(2+)/calmodulin metabolism, also increased, while downregulated genes included several ion-channel components, such as Fxyd2. Bioinformatics analyses suggest that genes modulated by betaine are involved in DNA replication, might counteract UV-induced processes, and include many targets of transcription factors associated with cell proliferation and differentiation. Our results indicate that betaine controls unique gene expression pathways in keratinocytes, including some involved in differentiation.

  19. Spontaneous Activity Characteristics of 3D “Optonets”

    Science.gov (United States)

    Marom, Anat; Shor, Erez; Levenberg, Shulamit; Shoham, Shy

    2017-01-01

    Sporadic spontaneous network activity emerges during early central nervous system (CNS) development and, as the number of neuronal connections rises, the maturing network displays diverse and complex activity, including various types of synchronized patterns. These activity patterns have major implications on both basic research and the study of neurological disorders, and their interplay with network morphology tightly correlates with developmental events such as neuronal differentiation, migration and establishment of neurotransmitter phenotypes. Although 2D neural cultures models have provided important insights into network activity patterns, these cultures fail to mimic the complex 3D architecture of natural CNS neural networks and its consequences on connectivity and activity. A 3D in-vitro model mimicking early network development while enabling cellular-resolution observations, could thus significantly advance our understanding of the activity characteristics in the developing CNS. Here, we longitudinally studied the spontaneous activity patterns of developing 3D in-vitro neural network “optonets,” an optically-accessible bioengineered CNS model with multiple cortex-like characteristics. Optonet activity was observed using the genetically encodable calcium indicator GCaMP6m and a 3D imaging solution based on a standard epi-fluorescence microscope equipped with a piezo-electric z-stage and image processing-based deconvolution. Our results show that activity patterns become more complex as the network matures, gradually exhibiting longer-duration events. This report characterizes the patterns over time, and discusses how environmental changes affect the activity patterns. The relatively high degree of similarity between the network's spontaneously generated activity patterns and the reported characteristics of in-vivo activity, suggests that this is a compelling model system for brain-in-a chip research. PMID:28119555

  20. 3D modeling for the generation of virtual heritage

    Directory of Open Access Journals (Sweden)

    Francisco Díaz Gómez

    2015-10-01

    Full Text Available The present article is focused on the generation of virtual 3D contents from cultural heritage. Its main structure is divided in two well-defined blocks: the first one focused in the generation of 3D models, analyzing the most used technologies of 3D measuring in the cultural heritage, the most important software applications for the management of the 3D models obtained and the generation of the target contents; and a second block for exposing two case studies showing potential of these technologies, previously shown, for approaching the cultural heritage to both the general public and researchers, due to the development of the information and communication technologies.

  1. The dimension added by 3D scanning and 3D printing of meteorites

    Science.gov (United States)

    de Vet, S. J.

    2016-01-01

    An overview for the 3D photodocumentation of meteorites is presented, focussing on two 3D scanning methods in relation to 3D printing. The 3D photodocumention of meteorites provides new ways for the digital preservation of culturally, historically or scientifically unique meteorites. It has the potential for becoming a new documentation standard of meteorites that can exist complementary to traditional photographic documentation. Notable applications include (i.) use of physical properties in dark flight-, strewn field-, or aerodynamic modelling; (ii.) collection research of meteorites curated by different museum collections, and (iii.) public dissemination of meteorite models as a resource for educational users. The possible applications provided by the additional dimension of 3D illustrate the benefits for the meteoritics community.

  2. From Mental Game to Cultural Praxis: A Cultural Studies Model's Implications for the Future of Sport Psychology

    Science.gov (United States)

    Ryba, Tatiana V.; Wright, Handel Kashope

    2005-01-01

    This paper explores the implications of a cultural studies as praxis heuristic "model: for transforming sport psychology". It provides a brief introduction to both cultural studies and sport psychology and discusses a cultural studies intersection with sport studies and sport psychology. Cultural studies, it asserts, provides one of several…

  3. Holography of 3d-3d correspondence at Large N

    OpenAIRE

    Gang, Dongmin; Kim, Nakwoo; Lee, Sangmin

    2014-01-01

    We study the physics of multiple M5-branes compactified on a hyperbolic 3-manifold. On the one hand, it leads to the 3d-3d correspondence which maps an N = 2 $$ \\mathcal{N}=2 $$ superconformal field theory to a pure Chern-Simons theory on the 3-manifold. On the other hand, it leads to a warped AdS 4 geometry in M-theory holographically dual to the superconformal field theory. Combining the holographic duality and the 3d-3d correspondence, we propose a conjecture for the large N limit of the p...

  4. EU Design Law and 3D Printing

    DEFF Research Database (Denmark)

    Nordberg, Ana; Schovsbo, Jens Hemmingsen

    2017-01-01

    The article considers the implications for EU design law of 3D-printing. It first describes the 3D-printing technology and the e-ecosystem which is evolving around the technology and involves a number of new stakeholders who in different ways are engaged in the making and sharing of CAD-files and....../or printing. It is submitted that it is only a matter of time before 3D-printing equipment becomes ubiquitous. It is pointed out how the new technology and e-ecosystem at the same time represent threats and opportunities to design holders and to the societal interests in design and design law. EU design law...

  5. [Culture and cultural gaps in work teams: implications for organisational commitment].

    Science.gov (United States)

    Sánchez, José C; Lanero, Ana; Yurrebaso, Amaia; Tejero, Blanca

    2007-05-01

    Some theoreticians of organisational commitment have proposed that culture is an important determinant of organisational commitment. Nevertheless, very few studies have examined the role that work teams culture (subculture) and their cultural gaps play in commitment. This study is an attempt to overcome this lack. Using a sample of 375 work teams from various public and private organisations, it was found that the results confirmed our proposals. Cultural gaps were negatively related to commitment; the teams subculture was positively related to commitment, and more highly to commitment to values than to commitment to continuing. Contrary to the results of other studies, the demographic variables (age, time on the team, time in the company) were not significant, except that educational level was related to the commitment to continue. The implications of these results are analysed.

  6. 3D Spectroscopy in Astronomy

    Science.gov (United States)

    Mediavilla, Evencio; Arribas, Santiago; Roth, Martin; Cepa-Nogué, Jordi; Sánchez, Francisco

    2011-09-01

    Preface; Acknowledgements; 1. Introductory review and technical approaches Martin M. Roth; 2. Observational procedures and data reduction James E. H. Turner; 3. 3D Spectroscopy instrumentation M. A. Bershady; 4. Analysis of 3D data Pierre Ferruit; 5. Science motivation for IFS and galactic studies F. Eisenhauer; 6. Extragalactic studies and future IFS science Luis Colina; 7. Tutorials: how to handle 3D spectroscopy data Sebastian F. Sánchez, Begona García-Lorenzo and Arlette Pécontal-Rousset.

  7. Spherical 3D isotropic wavelets

    Science.gov (United States)

    Lanusse, F.; Rassat, A.; Starck, J.-L.

    2012-04-01

    Context. Future cosmological surveys will provide 3D large scale structure maps with large sky coverage, for which a 3D spherical Fourier-Bessel (SFB) analysis in spherical coordinates is natural. Wavelets are particularly well-suited to the analysis and denoising of cosmological data, but a spherical 3D isotropic wavelet transform does not currently exist to analyse spherical 3D data. Aims: The aim of this paper is to present a new formalism for a spherical 3D isotropic wavelet, i.e. one based on the SFB decomposition of a 3D field and accompany the formalism with a public code to perform wavelet transforms. Methods: We describe a new 3D isotropic spherical wavelet decomposition based on the undecimated wavelet transform (UWT) described in Starck et al. (2006). We also present a new fast discrete spherical Fourier-Bessel transform (DSFBT) based on both a discrete Bessel transform and the HEALPIX angular pixelisation scheme. We test the 3D wavelet transform and as a toy-application, apply a denoising algorithm in wavelet space to the Virgo large box cosmological simulations and find we can successfully remove noise without much loss to the large scale structure. Results: We have described a new spherical 3D isotropic wavelet transform, ideally suited to analyse and denoise future 3D spherical cosmological surveys, which uses a novel DSFBT. We illustrate its potential use for denoising using a toy model. All the algorithms presented in this paper are available for download as a public code called MRS3D at http://jstarck.free.fr/mrs3d.html

  8. Xenobiotic metabolism capacities of human skin in comparison with a 3D-epidermis model and keratinocyte-based cell culture as in vitro alternatives for chemical testing: phase II enzymes.

    Science.gov (United States)

    Götz, Christine; Pfeiffer, Roland; Tigges, Julia; Ruwiedel, Karsten; Hübenthal, Ulrike; Merk, Hans F; Krutmann, Jean; Edwards, Robert J; Abel, Josef; Pease, Camilla; Goebel, Carsten; Hewitt, Nicola; Fritsche, Ellen

    2012-05-01

    The 7th Amendment to the EU Cosmetics Directive prohibits the use of animals in cosmetic testing for certain endpoints, such as genotoxicity. Therefore, skin in vitro models have to replace chemical testing in vivo. However, the metabolic competence neither of human skin nor of alternative in vitro models has so far been fully characterized, although skin is the first-pass organ for accidentally or purposely (cosmetics and pharmaceuticals) applied chemicals. Thus, there is an urgent need to understand the xenobiotic-metabolizing capacities of human skin and to compare these activities to models developed to replace animal testing. We have measured the activity of the phase II enzymes glutathione S-transferase, UDP-glucuronosyltransferase and N-acetyltransferase in ex vivo human skin, the 3D epidermal model EpiDerm 200 (EPI-200), immortalized keratinocyte-based cell lines (HaCaT and NCTC 2544) and primary normal human epidermal keratinocytes. We show that all three phase II enzymes are present and highly active in skin as compared to phase I. Human skin, therefore, represents a more detoxifying than activating organ. This work systematically compares the activities of three important phase II enzymes in four different in vitro models directly to human skin. We conclude from our studies that 3D epidermal models, like the EPI-200 employed here, are superior over monolayer cultures in mimicking human skin xenobiotic metabolism and thus better suited for dermatotoxicity testing. © 2012 John Wiley & Sons A/S.

  9. 3D IBFV : Hardware-Accelerated 3D Flow Visualization

    NARCIS (Netherlands)

    Telea, Alexandru; Wijk, Jarke J. van

    2003-01-01

    We present a hardware-accelerated method for visualizing 3D flow fields. The method is based on insertion, advection, and decay of dye. To this aim, we extend the texture-based IBFV technique for 2D flow visualization in two main directions. First, we decompose the 3D flow visualization problem in a

  10. 3D Elevation Program—Virtual USA in 3D

    Science.gov (United States)

    Lukas, Vicki; Stoker, J.M.

    2016-04-14

    The U.S. Geological Survey (USGS) 3D Elevation Program (3DEP) uses a laser system called ‘lidar’ (light detection and ranging) to create a virtual reality map of the Nation that is very accurate. 3D maps have many uses with new uses being discovered all the time.  

  11. A 3-D Contextual Classifier

    DEFF Research Database (Denmark)

    Larsen, Rasmus

    1997-01-01

    . This includes the specification of a Gaussian distribution for the pixel values as well as a prior distribution for the configuration of class variables within the cross that is m ade of a pixel and its four nearest neighbours. We will extend this algorithm to 3-D, i.e. we will specify a simultaneous Gaussian...... distr ibution for a pixel and its 6 nearest 3-D neighbours, and generalise the class variable configuration distribution within the 3-D cross. The algorithm is tested on a synthetic 3-D multivariate dataset....

  12. 3D Bayesian contextual classifiers

    DEFF Research Database (Denmark)

    Larsen, Rasmus

    2000-01-01

    We extend a series of multivariate Bayesian 2-D contextual classifiers to 3-D by specifying a simultaneous Gaussian distribution for the feature vectors as well as a prior distribution of the class variables of a pixel and its 6 nearest 3-D neighbours.......We extend a series of multivariate Bayesian 2-D contextual classifiers to 3-D by specifying a simultaneous Gaussian distribution for the feature vectors as well as a prior distribution of the class variables of a pixel and its 6 nearest 3-D neighbours....

  13. Using 3D in Visualization

    DEFF Research Database (Denmark)

    Wood, Jo; Kirschenbauer, Sabine; Döllner, Jürgen

    2005-01-01

    to display 3D imagery. The extra cartographic degree of freedom offered by using 3D is explored and offered as a motivation for employing 3D in visualization. The use of VR and the construction of virtual environments exploit navigational and behavioral realism, but become most usefil when combined...... with abstracted representations embedded in a 3D space. The interactions between development of geovisualization, the technology used to implement it and the theory surrounding cartographic representation are explored. The dominance of computing technologies, driven particularly by the gaming industry...

  14. Interactive 3D multimedia content

    CERN Document Server

    Cellary, Wojciech

    2012-01-01

    The book describes recent research results in the areas of modelling, creation, management and presentation of interactive 3D multimedia content. The book describes the current state of the art in the field and identifies the most important research and design issues. Consecutive chapters address these issues. These are: database modelling of 3D content, security in 3D environments, describing interactivity of content, searching content, visualization of search results, modelling mixed reality content, and efficient creation of interactive 3D content. Each chapter is illustrated with example a

  15. 3D for Graphic Designers

    CERN Document Server

    Connell, Ellery

    2011-01-01

    Helping graphic designers expand their 2D skills into the 3D space The trend in graphic design is towards 3D, with the demand for motion graphics, animation, photorealism, and interactivity rapidly increasing. And with the meteoric rise of iPads, smartphones, and other interactive devices, the design landscape is changing faster than ever.2D digital artists who need a quick and efficient way to join this brave new world will want 3D for Graphic Designers. Readers get hands-on basic training in working in the 3D space, including product design, industrial design and visualization, modeling, ani

  16. 3-D printers for libraries

    CERN Document Server

    Griffey, Jason

    2014-01-01

    As the maker movement continues to grow and 3-D printers become more affordable, an expanding group of hobbyists is keen to explore this new technology. In the time-honored tradition of introducing new technologies, many libraries are considering purchasing a 3-D printer. Jason Griffey, an early enthusiast of 3-D printing, has researched the marketplace and seen several systems first hand at the Consumer Electronics Show. In this report he introduces readers to the 3-D printing marketplace, covering such topics asHow fused deposition modeling (FDM) printing workBasic terminology such as build

  17. Perception of 3D spatial relations for 3D displays

    Science.gov (United States)

    Rosen, Paul; Pizlo, Zygmunt; Hoffmann, Christoph; Popescu, Voicu S.

    2004-05-01

    We test perception of 3D spatial relations in 3D images rendered by a 3D display (Perspecta from Actuality Systems) and compare it to that of a high-resolution flat panel display. 3D images provide the observer with such depth cues as motion parallax and binocular disparity. Our 3D display is a device that renders a 3D image by displaying, in rapid succession, radial slices through the scene on a rotating screen. The image is contained in a glass globe and can be viewed from virtually any direction. In the psychophysical experiment several families of 3D objects are used as stimuli: primitive shapes (cylinders and cuboids), and complex objects (multi-story buildings, cars, and pieces of furniture). Each object has at least one plane of symmetry. On each trial an object or its "distorted" version is shown at an arbitrary orientation. The distortion is produced by stretching an object in a random direction by 40%. This distortion must eliminate the symmetry of an object. The subject's task is to decide whether or not the presented object is distorted under several viewing conditions (monocular/binocular, with/without motion parallax, and near/far). The subject's performance is measured by the discriminability d', which is a conventional dependent variable in signal detection experiments.

  18. 3D Printing for Bricks

    OpenAIRE

    ECT Team, Purdue

    2015-01-01

    Building Bytes, by Brian Peters, is a project that uses desktop 3D printers to print bricks for architecture. Instead of using an expensive custom-made printer, it uses a normal standard 3D printer which is available for everyone and makes it more accessible and also easier for fabrication.

  19. Market study: 3-D eyetracker

    Science.gov (United States)

    1977-01-01

    A market study of a proposed version of a 3-D eyetracker for initial use at NASA's Ames Research Center was made. The commercialization potential of a simplified, less expensive 3-D eyetracker was ascertained. Primary focus on present and potential users of eyetrackers, as well as present and potential manufacturers has provided an effective means of analyzing the prospects for commercialization.

  20. Spherical 3D Isotropic Wavelets

    CERN Document Server

    Lanusse, F; Starck, J -L

    2011-01-01

    Future cosmological surveys will provide 3D large scale structure maps with large sky coverage, for which a 3D Spherical Fourier-Bessel (SFB) analysis in is natural. Wavelets are particularly well-suited to the analysis and denoising of cosmological data, but a spherical 3D isotropic wavelet transform does not currently exist to analyse spherical 3D data. The aim of this paper is to present a new formalism for a spherical 3D isotropic wavelet, i.e. one based on the Fourier-Bessel decomposition of a 3D field and accompany the formalism with a public code to perform wavelet transforms. We describe a new 3D isotropic spherical wavelet decomposition based on the undecimated wavelet transform (UWT) described in Starck et al. 2006. We also present a new fast Discrete Spherical Fourier-Bessel Transform (DSFBT) based on both a discrete Bessel Transform and the HEALPIX angular pixelisation scheme. We test the 3D wavelet transform and as a toy-application, apply a denoising algorithm in wavelet space to the Virgo large...

  1. Improvement of 3D Scanner

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    The disadvantage remaining in 3D scanning system and its reasons are discussed. A new host-and-slave structure with high speed image acquisition and processing system is proposed to quicken the image processing and improve the performance of 3D scanning system.

  2. Advanced 3-D Ultrasound Imaging

    DEFF Research Database (Denmark)

    Rasmussen, Morten Fischer

    to produce high quality 3-D images. Because of the large matrix transducers with integrated custom electronics, these systems are extremely expensive. The relatively low price of ultrasound scanners is one of the factors for the widespread use of ultrasound imaging. The high price tag on the high quality 3-D......The main purpose of the PhD project was to develop methods that increase the 3-D ultrasound imaging quality available for the medical personnel in the clinic. Acquiring a 3-D volume gives the medical doctor the freedom to investigate the measured anatomy in any slice desirable after the scan has...... been completed. This allows for precise measurements of organs dimensions and makes the scan more operator independent. Real-time 3-D ultrasound imaging is still not as widespread in use in the clinics as 2-D imaging. A limiting factor has traditionally been the low image quality achievable using...

  3. Using 3D in Visualization

    DEFF Research Database (Denmark)

    Wood, Jo; Kirschenbauer, Sabine; Döllner, Jürgen

    2005-01-01

    The notion of three-dimensionality is applied to five stages of the visualization pipeline. While 3D visulization is most often associated with the visual mapping and representation of data, this chapter also identifies its role in the management and assembly of data, and in the media used...... to display 3D imagery. The extra cartographic degree of freedom offered by using 3D is explored and offered as a motivation for employing 3D in visualization. The use of VR and the construction of virtual environments exploit navigational and behavioral realism, but become most usefil when combined...... with abstracted representations embedded in a 3D space. The interactions between development of geovisualization, the technology used to implement it and the theory surrounding cartographic representation are explored. The dominance of computing technologies, driven particularly by the gaming industry...

  4. 3D printing in dentistry.

    Science.gov (United States)

    Dawood, A; Marti Marti, B; Sauret-Jackson, V; Darwood, A

    2015-12-01

    3D printing has been hailed as a disruptive technology which will change manufacturing. Used in aerospace, defence, art and design, 3D printing is becoming a subject of great interest in surgery. The technology has a particular resonance with dentistry, and with advances in 3D imaging and modelling technologies such as cone beam computed tomography and intraoral scanning, and with the relatively long history of the use of CAD CAM technologies in dentistry, it will become of increasing importance. Uses of 3D printing include the production of drill guides for dental implants, the production of physical models for prosthodontics, orthodontics and surgery, the manufacture of dental, craniomaxillofacial and orthopaedic implants, and the fabrication of copings and frameworks for implant and dental restorations. This paper reviews the types of 3D printing technologies available and their various applications in dentistry and in maxillofacial surgery.

  5. 3D vision system assessment

    Science.gov (United States)

    Pezzaniti, J. Larry; Edmondson, Richard; Vaden, Justin; Hyatt, Bryan; Chenault, David B.; Kingston, David; Geulen, Vanilynmae; Newell, Scott; Pettijohn, Brad

    2009-02-01

    In this paper, we report on the development of a 3D vision system consisting of a flat panel stereoscopic display and auto-converging stereo camera and an assessment of the system's use for robotic driving, manipulation, and surveillance operations. The 3D vision system was integrated onto a Talon Robot and Operator Control Unit (OCU) such that direct comparisons of the performance of a number of test subjects using 2D and 3D vision systems were possible. A number of representative scenarios were developed to determine which tasks benefited most from the added depth perception and to understand when the 3D vision system hindered understanding of the scene. Two tests were conducted at Fort Leonard Wood, MO with noncommissioned officers ranked Staff Sergeant and Sergeant First Class. The scenarios; the test planning, approach and protocols; the data analysis; and the resulting performance assessment of the 3D vision system are reported.

  6. PLOT3D user's manual

    Science.gov (United States)

    Walatka, Pamela P.; Buning, Pieter G.; Pierce, Larry; Elson, Patricia A.

    1990-01-01

    PLOT3D is a computer graphics program designed to visualize the grids and solutions of computational fluid dynamics. Seventy-four functions are available. Versions are available for many systems. PLOT3D can handle multiple grids with a million or more grid points, and can produce varieties of model renderings, such as wireframe or flat shaded. Output from PLOT3D can be used in animation programs. The first part of this manual is a tutorial that takes the reader, keystroke by keystroke, through a PLOT3D session. The second part of the manual contains reference chapters, including the helpfile, data file formats, advice on changing PLOT3D, and sample command files.

  7. ADT-3D Tumor Detection Assistant in 3D

    Directory of Open Access Journals (Sweden)

    Jaime Lazcano Bello

    2008-12-01

    Full Text Available The present document describes ADT-3D (Three-Dimensional Tumor Detector Assistant, a prototype application developed to assist doctors diagnose, detect and locate tumors in the brain by using CT scan. The reader may find on this document an introduction to tumor detection; ADT-3D main goals; development details; description of the product; motivation for its development; result’s study; and areas of applicability.

  8. Unassisted 3D camera calibration

    Science.gov (United States)

    Atanassov, Kalin; Ramachandra, Vikas; Nash, James; Goma, Sergio R.

    2012-03-01

    With the rapid growth of 3D technology, 3D image capture has become a critical part of the 3D feature set on mobile phones. 3D image quality is affected by the scene geometry as well as on-the-device processing. An automatic 3D system usually assumes known camera poses accomplished by factory calibration using a special chart. In real life settings, pose parameters estimated by factory calibration can be negatively impacted by movements of the lens barrel due to shaking, focusing, or camera drop. If any of these factors displaces the optical axes of either or both cameras, vertical disparity might exceed the maximum tolerable margin and the 3D user may experience eye strain or headaches. To make 3D capture more practical, one needs to consider unassisted (on arbitrary scenes) calibration. In this paper, we propose an algorithm that relies on detection and matching of keypoints between left and right images. Frames containing erroneous matches, along with frames with insufficiently rich keypoint constellations, are detected and discarded. Roll, pitch yaw , and scale differences between left and right frames are then estimated. The algorithm performance is evaluated in terms of the remaining vertical disparity as compared to the maximum tolerable vertical disparity.

  9. 3D GEO: AN ALTERNATIVE APPROACH

    Directory of Open Access Journals (Sweden)

    A. Georgopoulos

    2016-10-01

    Full Text Available The expression GEO is mostly used to denote relation to the earth. However it should not be confined to what is related to the earth's surface, as other objects also need three dimensional representation and documentation, like cultural heritage objects. They include both tangible and intangible ones. In this paper the 3D data acquisition and 3D modelling of cultural heritage assets are briefly described and their significance is also highlighted. Moreover the organization of such information, related to monuments and artefacts, into relational data bases and its use for various purposes, other than just geometric documentation is also described and presented. In order to help the reader understand the above, several characteristic examples are presented and their methodology explained and their results evaluated.

  10. 3D Geo: An Alternative Approach

    Science.gov (United States)

    Georgopoulos, A.

    2016-10-01

    The expression GEO is mostly used to denote relation to the earth. However it should not be confined to what is related to the earth's surface, as other objects also need three dimensional representation and documentation, like cultural heritage objects. They include both tangible and intangible ones. In this paper the 3D data acquisition and 3D modelling of cultural heritage assets are briefly described and their significance is also highlighted. Moreover the organization of such information, related to monuments and artefacts, into relational data bases and its use for various purposes, other than just geometric documentation is also described and presented. In order to help the reader understand the above, several characteristic examples are presented and their methodology explained and their results evaluated.

  11. Direct inoculation method using BacT/ALERT 3D and BD Phoenix System allows rapid and accurate identification and susceptibility testing for both Gram-positive cocci and Gram-negative rods in aerobic blood cultures.

    Science.gov (United States)

    Yonetani, Shota; Okazaki, Mitsuhiro; Araki, Koji; Makino, Hiroshi; Fukugawa, Yoko; Okuyama, Takahiro; Ohnishi, Hiroaki; Watanabe, Takashi

    2012-06-01

    This study describes a direct inoculation method using the automated BacT/ALERT 3D and the BD Phoenix System in combination for identification and susceptibility testing of isolates from positive blood cultures. Organism identification and susceptibility results were compared with the conventional method for 211 positive aerobic blood cultures. Of 110 Gram-positive cocci (GPCs), 98 (89.1%) isolates were correctly identified to the species level. Of 101 Gram-negative rods (GNRs), 98 (97.0%) isolates were correctly identified to the species level. The overall categorical agreement in antimicrobial susceptibility testing among the 110 GPCs was 92.7%, with 0.04% very major and 0.7% major error rates. The overall categorical agreement among 78 isolates of enterobacteria and 23 isolates of nonfermenters in GNRs was 99.5% and 91.1%, respectively, with no major errors identified. We conclude that, compared with previously reported direct inoculation methods, our method is superior in identification and susceptibility testing of GPCs.

  12. Tuotekehitysprojekti: 3D-tulostin

    OpenAIRE

    Pihlajamäki, Janne

    2011-01-01

    Opinnäytetyössä tutustuttiin 3D-tulostamisen teknologiaan. Työssä käytiin läpi 3D-tulostimesta tehty tuotekehitysprojekti. Sen lisäksi esiteltiin yleisellä tasolla tuotekehitysprosessi ja syntyneiden tulosten mahdollisia suojausmenetelmiä. Tavoitteena tässä työssä oli kehittää markkinoilta jo löytyvää kotitulostin-tasoista 3D-laiteteknologiaa lähemmäksi ammattilaistason ratkaisua. Tavoitteeseen pyrittiin keskittymällä parantamaan laitteella saavutettavaa tulostustarkkuutta ja -nopeutt...

  13. Color 3D Reverse Engineering

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    This paper presents a principle and a method of col or 3D laser scanning measurement. Based on the fundamental monochrome 3D measureme nt study, color information capture, color texture mapping, coordinate computati on and other techniques are performed to achieve color 3D measurement. The syste m is designed and composed of a line laser light emitter, one color CCD camera, a motor-driven rotary filter, a circuit card and a computer. Two steps in captu ring object's images in the measurement process: Firs...

  14. Exploration of 3D Printing

    OpenAIRE

    Lin, Zeyu

    2014-01-01

    3D printing technology is introduced and defined in this Thesis. Some methods of 3D printing are illustrated and their principles are explained with pictures. Most of the essential parts are presented with pictures and their effects are explained within the whole system. Problems on Up! Plus 3D printer are solved and a DIY product is made with this machine. The processes of making product are recorded and the items which need to be noticed during the process are the highlight in this th...

  15. Handbook of 3D integration

    CERN Document Server

    Garrou , Philip; Ramm , Peter

    2014-01-01

    Edited by key figures in 3D integration and written by top authors from high-tech companies and renowned research institutions, this book covers the intricate details of 3D process technology.As such, the main focus is on silicon via formation, bonding and debonding, thinning, via reveal and backside processing, both from a technological and a materials science perspective. The last part of the book is concerned with assessing and enhancing the reliability of the 3D integrated devices, which is a prerequisite for the large-scale implementation of this emerging technology. Invaluable reading fo

  16. 3D-ICONS Ireland – fulfilling the potential of a rich 3D resource

    Directory of Open Access Journals (Sweden)

    Anthony Corns

    2017-03-01

    Full Text Available As a partner in the EU co-funded 3D-ICONS project, the Discovery Programme undertook the 3D documentation of some of the most iconic cultural heritage sites in Ireland. This pan-European project aimed to establish a complete pipeline for the production of 3D replicas of archaeological monuments and historic buildings, and to publish the content to Europeana for public access. The list of Irish icons range from wider cultural landscapes to smaller ornately carved stones and includes a wide range of chronological periods: from Neolithic rock art from 2500 BC to Derry's 17th-century fortifications. The primary digitisation methods include airborne laser scanning (ALS, phase-based terrestrial laser scanning (Faro Focus 3D and close range structured light scanning (Artec EVA. These are now mainstream approaches for surveying historic landscapes, structures and objects, generating precise, high-resolution point cloud data, primarily for viewing and interaction in proprietary software applications. The challenge was to convert these complex high-volume datasets into textured 3D models, retaining the geometric integrity of the original data. The article highlights the development of a pipeline to produce a lightweight 3D model that enables the public to interact with a photorealistic model based upon accurate survey and texture data. 3D-ICONS ended in January 2015, but a new website 3dicons.ie was launched to offer continued access to the Irish 3D models and associated content and media generated during the project. The article will consider the impact of this online content, particularly how it has been used as a teaching aid in secondary schools and how this may be extended in the future. It will also demonstrate how content from the project has been remodelled to develop an interactive and immersive experience for the great mound at Knowth, a development in partnership with the operators of the Brú na Bóinne visitor centre.

  17. 3D VIRTUAL RECONSTRUCTION OF ARCHAEOLOGICAL MONUMENTS

    OpenAIRE

    2014-01-01

    3D Virtual Models are the future of the representation of the existing and destroyed architectural heritage. The term reconstruction defines the re-building of a monument to its state at the time of its history chosen for that particular representation. In recent years the evolution of the technology, has contributed significantly in many aspects of the field of cultural heritage preservation and recording. Techniques like digital image processing, digital orthophoto production, terrestrial l...

  18. Conducting polymer 3D microelectrodes

    DEFF Research Database (Denmark)

    Sasso, Luigi; Vazquez, Patricia; Vedarethinam, Indumathi

    2010-01-01

    Conducting polymer 3D microelectrodes have been fabricated for possible future neurological applications. A combination of micro-fabrication techniques and chemical polymerization methods has been used to create pillar electrodes in polyaniline and polypyrrole. The thin polymer films obtained...

  19. Accepting the T3D

    Energy Technology Data Exchange (ETDEWEB)

    Rich, D.O.; Pope, S.C.; DeLapp, J.G.

    1994-10-01

    In April, a 128 PE Cray T3D was installed at Los Alamos National Laboratory`s Advanced Computing Laboratory as part of the DOE`s High-Performance Parallel Processor Program (H4P). In conjunction with CRI, the authors implemented a 30 day acceptance test. The test was constructed in part to help them understand the strengths and weaknesses of the T3D. In this paper, they briefly describe the H4P and its goals. They discuss the design and implementation of the T3D acceptance test and detail issues that arose during the test. They conclude with a set of system requirements that must be addressed as the T3D system evolves.

  20. 3-D Vector Flow Imaging

    DEFF Research Database (Denmark)

    Holbek, Simon

    For the last decade, the field of ultrasonic vector flow imaging has gotten an increasingly attention, as the technique offers a variety of new applications for screening and diagnostics of cardiovascular pathologies. The main purpose of this PhD project was therefore to advance the field of 3-D...... ultrasonic vector flow estimation and bring it a step closer to a clinical application. A method for high frame rate 3-D vector flow estimation in a plane using the transverse oscillation method combined with a 1024 channel 2-D matrix array is presented. The proposed method is validated both through phantom......, if this significant reduction in the element count can still provide precise and robust 3-D vector flow estimates in a plane. The study concludes that the RC array is capable of estimating precise 3-D vector flow both in a plane and in a volume, despite the low channel count. However, some inherent new challenges...

  1. 3D Face Apperance Model

    DEFF Research Database (Denmark)

    Lading, Brian; Larsen, Rasmus; Astrom, K

    2006-01-01

    We build a 3D face shape model, including inter- and intra-shape variations, derive the analytical Jacobian of its resulting 2D rendered image, and show example of its fitting performance with light, pose, id, expression and texture variations......We build a 3D face shape model, including inter- and intra-shape variations, derive the analytical Jacobian of its resulting 2D rendered image, and show example of its fitting performance with light, pose, id, expression and texture variations...

  2. 3D Face Appearance Model

    DEFF Research Database (Denmark)

    Lading, Brian; Larsen, Rasmus; Åström, Kalle

    2006-01-01

    We build a 3d face shape model, including inter- and intra-shape variations, derive the analytical jacobian of its resulting 2d rendered image, and show example of its fitting performance with light, pose, id, expression and texture variations.}......We build a 3d face shape model, including inter- and intra-shape variations, derive the analytical jacobian of its resulting 2d rendered image, and show example of its fitting performance with light, pose, id, expression and texture variations.}...

  3. Main: TATCCAYMOTIFOSRAMY3D [PLACE

    Lifescience Database Archive (English)

    Full Text Available TATCCAYMOTIFOSRAMY3D S000256 01-August-2006 (last modified) kehi TATCCAY motif foun...d in rice (O.s.) RAmy3D alpha-amylase gene promoter; Y=T/C; a GATA motif as its antisense sequence; TATCCAY ...motif and G motif (see S000130) are responsible for sugar repression (Toyofuku et al. 1998); GATA; amylase; sugar; repression; rice (Oryza sativa) TATCCAY ...

  4. Cosmic evolution: the context for astrobiology and its cultural implications

    Science.gov (United States)

    Dick, Steven J.

    2012-10-01

    Astrobiology must be seen in the context of cosmic evolution, the 13.7 billion-year master narrative of the universe. The idea of an evolving universe dates back only to the 19th century, and became a guiding principle for astronomical research only in the second half of the 20th century. The modern synthesis in evolutionary biology hastened the acceptance of the idea in its cosmic setting, as did the confirmation of the Big Bang theory for the origin of the universe. NASA programmes such as Origins incorporated it as a guiding principle. Cosmic evolution encompasses physical, biological and cultural evolution, and may result in a physical, biological or postbiological universe, each with its own implications for long-term human destiny, and each imbuing the meaning of life with different values. It has the status of an increasingly accepted worldview that is beginning to have a profound effect not only in science but also in religion and philosophy.

  5. MPML3D: Scripting Agents for the 3D Internet.

    Science.gov (United States)

    Prendinger, Helmut; Ullrich, Sebastian; Nakasone, Arturo; Ishizuka, Mitsuru

    2011-05-01

    The aim of this paper is two-fold. First, it describes a scripting language for specifying communicative behavior and interaction of computer-controlled agents ("bots") in the popular three-dimensional (3D) multiuser online world of "Second Life" and the emerging "OpenSimulator" project. While tools for designing avatars and in-world objects in Second Life exist, technology for nonprogrammer content creators of scenarios involving scripted agents is currently missing. Therefore, we have implemented new client software that controls bots based on the Multimodal Presentation Markup Language 3D (MPML3D), a highly expressive XML-based scripting language for controlling the verbal and nonverbal behavior of interacting animated agents. Second, the paper compares Second Life and OpenSimulator platforms and discusses the merits and limitations of each from the perspective of agent control. Here, we also conducted a small study that compares the network performance of both platforms.

  6. Segmentation of Image Data from Complex Organotypic 3D Models of Cancer Tissues with Markov Random Fields

    Science.gov (United States)

    Robinson, Sean; Guyon, Laurent; Nevalainen, Jaakko; Toriseva, Mervi

    2015-01-01

    Organotypic, three dimensional (3D) cell culture models of epithelial tumour types such as prostate cancer recapitulate key aspects of the architecture and histology of solid cancers. Morphometric analysis of multicellular 3D organoids is particularly important when additional components such as the extracellular matrix and tumour microenvironment are included in the model. The complexity of such models has so far limited their successful implementation. There is a great need for automatic, accurate and robust image segmentation tools to facilitate the analysis of such biologically relevant 3D cell culture models. We present a segmentation method based on Markov random fields (MRFs) and illustrate our method using 3D stack image data from an organotypic 3D model of prostate cancer cells co-cultured with cancer-associated fibroblasts (CAFs). The 3D segmentation output suggests that these cell types are in physical contact with each other within the model, which has important implications for tumour biology. Segmentation performance is quantified using ground truth labels and we show how each step of our method increases segmentation accuracy. We provide the ground truth labels along with the image data and code. Using independent image data we show that our segmentation method is also more generally applicable to other types of cellular microscopy and not only limited to fluorescence microscopy. PMID:26630674

  7. Segmentation of Image Data from Complex Organotypic 3D Models of Cancer Tissues with Markov Random Fields.

    Directory of Open Access Journals (Sweden)

    Sean Robinson

    Full Text Available Organotypic, three dimensional (3D cell culture models of epithelial tumour types such as prostate cancer recapitulate key aspects of the architecture and histology of solid cancers. Morphometric analysis of multicellular 3D organoids is particularly important when additional components such as the extracellular matrix and tumour microenvironment are included in the model. The complexity of such models has so far limited their successful implementation. There is a great need for automatic, accurate and robust image segmentation tools to facilitate the analysis of such biologically relevant 3D cell culture models. We present a segmentation method based on Markov random fields (MRFs and illustrate our method using 3D stack image data from an organotypic 3D model of prostate cancer cells co-cultured with cancer-associated fibroblasts (CAFs. The 3D segmentation output suggests that these cell types are in physical contact with each other within the model, which has important implications for tumour biology. Segmentation performance is quantified using ground truth labels and we show how each step of our method increases segmentation accuracy. We provide the ground truth labels along with the image data and code. Using independent image data we show that our segmentation method is also more generally applicable to other types of cellular microscopy and not only limited to fluorescence microscopy.

  8. 3D-mallinnus ja 3D-animaatiot biovoimalaitoksesta

    OpenAIRE

    Hiltula, Tytti

    2014-01-01

    Opinnäytetyössä tehtiin biovoimalaitoksen piirustuksista 3D-mallinnus ja animaatiot. Työn tarkoituksena oli saada valmiiksi Recwell Oy:lle markkinointiin tarkoitetut kuva- ja videomateriaalit. Työssä perehdyttiin 3D-mallintamisen perustietoihin ja lähtökohtiin sekä animaation laatimiseen. Työ laadittiin kokonaisuudessaan AutoCAD-ohjelmalla, ja työn aikana tutustuttiin huolellisesti myös ohjelman käyttöohjeisiin. Piirustusten mitoituksessa huomattiin jo alkuvaiheessa suuria puutteita, ...

  9. From 3D view to 3D print

    Science.gov (United States)

    Dima, M.; Farisato, G.; Bergomi, M.; Viotto, V.; Magrin, D.; Greggio, D.; Farinato, J.; Marafatto, L.; Ragazzoni, R.; Piazza, D.

    2014-08-01

    In the last few years 3D printing is getting more and more popular and used in many fields going from manufacturing to industrial design, architecture, medical support and aerospace. 3D printing is an evolution of bi-dimensional printing, which allows to obtain a solid object from a 3D model, realized with a 3D modelling software. The final product is obtained using an additive process, in which successive layers of material are laid down one over the other. A 3D printer allows to realize, in a simple way, very complex shapes, which would be quite difficult to be produced with dedicated conventional facilities. Thanks to the fact that the 3D printing is obtained superposing one layer to the others, it doesn't need any particular work flow and it is sufficient to simply draw the model and send it to print. Many different kinds of 3D printers exist based on the technology and material used for layer deposition. A common material used by the toner is ABS plastics, which is a light and rigid thermoplastic polymer, whose peculiar mechanical properties make it diffusely used in several fields, like pipes production and cars interiors manufacturing. I used this technology to create a 1:1 scale model of the telescope which is the hardware core of the space small mission CHEOPS (CHaracterising ExOPlanets Satellite) by ESA, which aims to characterize EXOplanets via transits observations. The telescope has a Ritchey-Chrétien configuration with a 30cm aperture and the launch is foreseen in 2017. In this paper, I present the different phases for the realization of such a model, focusing onto pros and cons of this kind of technology. For example, because of the finite printable volume (10×10×12 inches in the x, y and z directions respectively), it has been necessary to split the largest parts of the instrument in smaller components to be then reassembled and post-processed. A further issue is the resolution of the printed material, which is expressed in terms of layers

  10. YouDash3D: exploring stereoscopic 3D gaming for 3D movie theaters

    Science.gov (United States)

    Schild, Jonas; Seele, Sven; Masuch, Maic

    2012-03-01

    Along with the success of the digitally revived stereoscopic cinema, events beyond 3D movies become attractive for movie theater operators, i.e. interactive 3D games. In this paper, we present a case that explores possible challenges and solutions for interactive 3D games to be played by a movie theater audience. We analyze the setting and showcase current issues related to lighting and interaction. Our second focus is to provide gameplay mechanics that make special use of stereoscopy, especially depth-based game design. Based on these results, we present YouDash3D, a game prototype that explores public stereoscopic gameplay in a reduced kiosk setup. It features live 3D HD video stream of a professional stereo camera rig rendered in a real-time game scene. We use the effect to place the stereoscopic effigies of players into the digital game. The game showcases how stereoscopic vision can provide for a novel depth-based game mechanic. Projected trigger zones and distributed clusters of the audience video allow for easy adaptation to larger audiences and 3D movie theater gaming.

  11. Materialedreven 3d digital formgivning

    DEFF Research Database (Denmark)

    Hansen, Flemming Tvede

    2010-01-01

    Formålet med forskningsprojektet er for det første at understøtte keramikeren i at arbejde eksperimenterende med digital formgivning, og for det andet at bidrage til en tværfaglig diskurs om brugen af digital formgivning. Forskningsprojektet fokuserer på 3d formgivning og derved på 3d digital...... formgivning og Rapid Prototyping (RP). RP er en fællesbetegnelse for en række af de teknikker, der muliggør at overføre den digitale form til 3d fysisk form. Forskningsprojektet koncentrerer sig om to overordnede forskningsspørgsmål. Det første handler om, hvordan viden og erfaring indenfor det keramiske...... fagområde kan blive udnyttet i forhold til 3d digital formgivning. Det andet handler om, hvad en sådan tilgang kan bidrage med, og hvordan den kan blive udnyttet i et dynamisk samspil med det keramiske materiale i formgivningen af 3d keramiske artefakter. Materialedreven formgivning er karakteriseret af en...

  12. Novel 3D media technologies

    CERN Document Server

    Dagiuklas, Tasos

    2015-01-01

    This book describes recent innovations in 3D media and technologies, with coverage of 3D media capturing, processing, encoding, and adaptation, networking aspects for 3D Media, and quality of user experience (QoE). The contributions are based on the results of the FP7 European Project ROMEO, which focuses on new methods for the compression and delivery of 3D multi-view video and spatial audio, as well as the optimization of networking and compression jointly across the future Internet. The delivery of 3D media to individual users remains a highly challenging problem due to the large amount of data involved, diverse network characteristics and user terminal requirements, as well as the user’s context such as their preferences and location. As the number of visual views increases, current systems will struggle to meet the demanding requirements in terms of delivery of consistent video quality to fixed and mobile users. ROMEO will present hybrid networking solutions that combine the DVB-T2 and DVB-NGH broadcas...

  13. 3D future internet media

    CERN Document Server

    Dagiuklas, Tasos

    2014-01-01

    This book describes recent innovations in 3D media and technologies, with coverage of 3D media capturing, processing, encoding, and adaptation, networking aspects for 3D Media, and quality of user experience (QoE). The main contributions are based on the results of the FP7 European Projects ROMEO, which focus on new methods for the compression and delivery of 3D multi-view video and spatial audio, as well as the optimization of networking and compression jointly across the Future Internet (www.ict-romeo.eu). The delivery of 3D media to individual users remains a highly challenging problem due to the large amount of data involved, diverse network characteristics and user terminal requirements, as well as the user’s context such as their preferences and location. As the number of visual views increases, current systems will struggle to meet the demanding requirements in terms of delivery of constant video quality to both fixed and mobile users. ROMEO will design and develop hybrid-networking solutions that co...

  14. Speaking Volumes About 3-D

    Science.gov (United States)

    2002-01-01

    In 1999, Genex submitted a proposal to Stennis Space Center for a volumetric 3-D display technique that would provide multiple users with a 360-degree perspective to simultaneously view and analyze 3-D data. The futuristic capabilities of the VolumeViewer(R) have offered tremendous benefits to commercial users in the fields of medicine and surgery, air traffic control, pilot training and education, computer-aided design/computer-aided manufacturing, and military/battlefield management. The technology has also helped NASA to better analyze and assess the various data collected by its satellite and spacecraft sensors. Genex capitalized on its success with Stennis by introducing two separate products to the commercial market that incorporate key elements of the 3-D display technology designed under an SBIR contract. The company Rainbow 3D(R) imaging camera is a novel, three-dimensional surface profile measurement system that can obtain a full-frame 3-D image in less than 1 second. The third product is the 360-degree OmniEye(R) video system. Ideal for intrusion detection, surveillance, and situation management, this unique camera system offers a continuous, panoramic view of a scene in real time.

  15. Modification of 3D milling machine to 3D printer

    OpenAIRE

    Halamíček, Lukáš

    2015-01-01

    Tato práce se zabývá přestavbou gravírovací frézky na 3D tiskárnu. V první části se práce zabývá možnými technologiemi 3D tisku a možností jejich využití u přestavby. Dále jsou popsány a vybrány vhodné součásti pro přestavbu. V další části je realizováno řízení ohřevu podložky, trysky a řízení posuvu drátu pomocí softwaru TwinCat od společnosti Beckhoff na průmyslovém počítači. Výsledkem práce by měla být oživená 3D tiskárna. This thesis deals with rebuilding of engraving machine to 3D pri...

  16. Aspects of defects in 3d-3d correspondence

    Energy Technology Data Exchange (ETDEWEB)

    Gang, Dongmin [Kavli Institute for the Physics and Mathematics of the Universe (WPI), University of Tokyo,Chiba 277-8583 (Japan); Kim, Nakwoo [Department of Physics and Research Institute of Basic Science, Kyung Hee University,Seoul 02447 (Korea, Republic of); School of Physics, Korea Institute for Advanced Study,Seoul 02455 (Korea, Republic of); Romo, Mauricio; Yamazaki, Masahito [Kavli Institute for the Physics and Mathematics of the Universe (WPI), University of Tokyo,Chiba 277-8583 (Japan); School of Natural Sciences, Institute for Advanced Study,Princeton, NJ 08540 (United States)

    2016-10-12

    In this paper we study supersymmetric co-dimension 2 and 4 defects in the compactification of the 6d (2,0) theory of type A{sub N−1} on a 3-manifold M. The so-called 3d-3d correspondence is a relation between complexified Chern-Simons theory (with gauge group SL(N,ℂ)) on M and a 3d N=2 theory T{sub N}[M]. We study this correspondence in the presence of supersymmetric defects, which are knots/links inside the 3-manifold. Our study employs a number of different methods: state-integral models for complex Chern-Simons theory, cluster algebra techniques, domain wall theory T[SU(N)], 5d N=2 SYM, and also supergravity analysis through holography. These methods are complementary and we find agreement between them. In some cases the results lead to highly non-trivial predictions on the partition function. Our discussion includes a general expression for the cluster partition function, which can be used to compute in the presence of maximal and certain class of non-maximal punctures when N>2. We also highlight the non-Abelian description of the 3d N=2T{sub N}[M] theory with defect included, when such a description is available. This paper is a companion to our shorter paper http://dx.doi.org/10.1088/1751-8113/49/30/30LT02, which summarizes our main results.

  17. Aspects of defects in 3d-3d correspondence

    Science.gov (United States)

    Gang, Dongmin; Kim, Nakwoo; Romo, Mauricio; Yamazaki, Masahito

    2016-10-01

    In this paper we study supersymmetric co-dimension 2 and 4 defects in the compactification of the 6d (2, 0) theory of type A N -1 on a 3-manifold M . The so-called 3d-3d correspondence is a relation between complexified Chern-Simons theory (with gauge group SL(N,C) ) on M and a 3d N=2 theory T N [ M ]. We study this correspondence in the presence of supersymmetric defects, which are knots/links inside the 3-manifold. Our study employs a number of different methods: state-integral models for complex Chern-Simons theory, cluster algebra techniques, domain wall theory T [SU( N )], 5d N=2 SYM, and also supergravity analysis through holography. These methods are complementary and we find agreement between them. In some cases the results lead to highly non-trivial predictions on the partition function. Our discussion includes a general expression for the cluster partition function, which can be used to compute in the presence of maximal and certain class of non-maximal punctures when N > 2. We also highlight the non-Abelian description of the 3d N=2 T N [ M ] theory with defect included, when such a description is available. This paper is a companion to our shorter paper [1], which summarizes our main results.

  18. Holography of 3d-3d correspondence at large N

    Energy Technology Data Exchange (ETDEWEB)

    Gang, Dongmin [School of Physics, Korea Institute for Advanced Study,85 Hoegiro, Dongdaemun-gu, Seoul, 130-722 (Korea, Republic of); Kim, Nakwoo [Department of Physics and Research Institute of Basic Sci