WorldWideScience

Sample records for 3d culture implications

  1. 3D PHOTOGRAPHS IN CULTURAL HERITAGE

    Schuhr, W.; J. D. Lee; Kiel, St.

    2013-01-01

    This paper on providing "oo-information" (= objective object-information) on cultural monuments and sites, based on 3D photographs is also a contribution of CIPA task group 3 to the 2013 CIPA Symposium in Strasbourg. To stimulate the interest in 3D photography for scientists as well as for amateurs, 3D-Masterpieces are presented. Exemplary it is shown, due to their high documentary value ("near reality"), 3D photography support, e.g. the recording, the visualization, the interpret...

  2. 3D Cell Culture in Alginate Hydrogels

    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. Filling gaps in cultural heritage documentation by 3D photography

    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.

  4. Multizone paper platform for 3D cell cultures.

    Ratmir Derda

    Full Text Available 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.

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

    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

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

    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

  7. EXPLORING CULTURAL HERITAGE RESOURCES IN A 3D COLLABORATIVE ENVIRONMENT

    A. Respaldiza; M. Wachowicz; Vázquez Hoehne, A.

    2012-01-01

    Cultural heritage is a complex and diverse concept, which brings together a wide domain of information. Resources linked to a cultural heritage site may consist of physical artefacts, books, works of art, pictures, historical maps, aerial photographs, archaeological surveys and 3D models. Moreover, all these resources are listed and described by a set of a variety of metadata specifications that allow their online search and consultation on the most basic characteristics of them. Some ex...

  8. Exploring Cultural Heritage Resources in a 3d Collaborative Environment

    Respaldiza, A.; Wachowicz, M.; Vázquez Hoehne, A.

    2012-06-01

    Cultural heritage is a complex and diverse concept, which brings together a wide domain of information. Resources linked to a cultural heritage site may consist of physical artefacts, books, works of art, pictures, historical maps, aerial photographs, archaeological surveys and 3D models. Moreover, all these resources are listed and described by a set of a variety of metadata specifications that allow their online search and consultation on the most basic characteristics of them. Some examples include Norma ISO 19115, Dublin Core, AAT, CDWA, CCO, DACS, MARC, MoReq, MODS, MuseumDat, TGN, SPECTRUM, VRA Core and Z39.50. Gateways are in place to fit in these metadata standards into those used in a SDI (ISO 19115 or INSPIRE), but substantial work still remains to be done for the complete incorporation of cultural heritage information. Therefore, the aim of this paper is to demonstrate how the complexity of cultural heritage resources can be dealt with by a visual exploration of their metadata within a 3D collaborative environment. The 3D collaborative environments are promising tools that represent the new frontier of our capacity of learning, understanding, communicating and transmitting culture.

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

    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.

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

    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.

  11. Surface modified alginate microcapsules for 3D cell culture

    Chen, Yi-Wen; Kuo, Chiung Wen; Chueh, Di-Yen; Chen, Peilin

    2016-06-01

    Culture as three dimensional cell aggregates or spheroids can offer an ideal platform for tissue engineering applications and for pharmaceutical screening. Such 3D culture models, however, may suffer from the problems such as immune response and ineffective and cumbersome culture. This paper describes a simple method for producing microcapsules with alginate cores and a thin shell of poly(L-lysine)-graft-poly(ethylene glycol) (PLL-g-PEG) to encapsulate mouse induced pluripotent stem (miPS) cells, generating a non-fouling surface as an effective immunoisolation barrier. We demonstrated the trapping of the alginate microcapsules in a microwell array for the continuous observation and culture of a large number of encapsulated miPS cells in parallel. miPS cells cultured in the microcapsules survived well and proliferated to form a single cell aggregate. Droplet formation of monodisperse microcapsules with controlled size combined with flow cytometry provided an efficient way to quantitatively analyze the growth of encapsulated cells in a high-throughput manner. The simple and cost-effective coating technique employed to produce the core-shell microcapsules could be used in the emerging field of cell therapy. The microwell array would provide a convenient, user friendly and high-throughput platform for long-term cell culture and monitoring.

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

    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. PMID:26164291

  13. Self-Assembled Peptide Gels for 3D Cell Culture

    Tang, Claire

    2010-01-01

    Under specific conditions short peptides modified with an N-terminal fluorenyl-9-methoxycarbonyl (Fmoc) group can self-assemble into hydrogel scaffolds similar in properties to the natural extracellular matrix. Fmoc-diphenylalanine (Fmoc-FF) for instance, has been shown to form hydrogels at physiological pH that have the ability to support 2D and 3D cell culture. The aim of this investigation is to provide further understanding of the self-assembly mechanism of such systems in order to progre...

  14. 3D visualization and virtual reality for cultural heritage diagnostic

    Lucio Colizzi

    2007-07-01

    Full Text Available In the past years, many different new technologies for Cultural Heritage Diagnostic have been developed. In particular laser scanner surveys with digital photogrammetry and also multi-spectral surveys are becoming very useful and inalienable tools for non invasive diagnosis. In the SIDART Project (Integrated System for Cultural Heritage diagnosis, we develop a software able to visualize and elaborate triangulated surfaces coming from high resolution laser scanner survey. In this paper, we want to present the most innovative aspect of our study, that is the possibility to visualize and work in default mode or in immersive Stereoscopy (3D mode. This lets the operator perceive the third dimension and the “virtual investigation” of the object becomes more realistic. This lets us take into consideration in a more simple, natural and correct way and also reduce the possibility to make wrong evaluation due to the false prospective of the classic visualization.

  15. Embedding Knowledge in 3D Data Frameworks in Cultural Heritage

    Coughenour, C. M.; Vincent, M. L.; de Kramer, M.; Senecal, S.; Fritsch, D.; Flores Gutirrez, M.; Lopez-Menchero Bendicho, V. M.; Ioannides, M.

    2015-08-01

    At present, where 3D modeling and visualisation in cultural heritage are concerned, an object's documentation lacks its interconnected memory provided by multidisciplinary examination and linked data. As the layers of paint, wood, and brick recount a structure's physical properties, the intangible, such as the forms of worship through song, dance, burning incense, and oral traditions, contributes to the greater story of its cultural heritage import. Furthermore, as an object or structure evolves through time, external political, religious, or environmental forces can affect it as well. As tangible and intangible entities associated with the structure transform, its narrative becomes dynamic and difficult to easily record. The Initial Training Network for Digital Cultural Heritage (ITN-DCH), a Marie Curie Actions project under the EU 7th Framework Programme, seeks to challenge this complexity by developing a novel methodology capable of offering such a holistic framework. With the integration of digitisation, conservation, linked data, and retrieval systems for DCH, the nature of investigation and dissemination will be augmented significantly. Examples of utilisating and evaluating this framework will range from a UNESCOWorld Heritage site, the Byzantine church of Panagia Forviotissa Asinou in the Troodos Mountains of Cyprus, to various religious icons and a monument located at the Monastery of Saint Neophytos. The application of this effort to the Asinou church, representing the first case study of the ITN-DCH project, is used as a template example in order to assess the technical challenges involved in the creation of such a framework.

  16. Diachronic 3d Reconstruction for Lost Cultural Heritage

    Guidi, G.; Russo, M.

    2011-09-01

    Cultural Heritage artifacts can often be underestimated for their hidden presence in the landscape. Such problem is particularly large in countries like Italy, where the massive amount of "famous" artifacts tends to neglect other presences unless properly exposed, or when the remains are dramatically damaged leaving very few interpretation clues to the visitor. In such cases a virtual presentation of the Cultural Heritage site can be of great help, specially for explaining the evolution of its status, giving sometimes sense to few spare stones. The definition of these digital representations deal with two crucial aspects: on the one hand the possibility of 3D surveying the relics in order to have an accurate geometrical image of the current status of the artifact; on the other hand the presence of historical sources both in form of written text or images, that once properly matched with the current geometrical data, may help to recreate digitally a set of 3D models representing visually the various historical phases (diachronic model), up to the current one. The core of this article is the definition of an integrated methodology that starts from an high-resolution digital survey of the remains of an ancient building and develops a coherent virtual reconstruction from different historical sources, suggesting a scalable method suitable to be re-used for generating a 4D (geometry + time) model of the artifact. This approach has been experimented on the "Basilica di San Giovanni in Conca" in Milan, a very significant example for its complex historic evolution that combines evident historic values with an invisible presence inside the city.

  17. DIACHRONIC 3D RECONSTRUCTION FOR LOST CULTURAL HERITAGE

    G. Guidi

    2012-09-01

    Full Text Available Cultural Heritage artifacts can often be underestimated for their hidden presence in the landscape. Such problem is particularly large in countries like Italy, where the massive amount of "famous" artifacts tends to neglect other presences unless properly exposed, or when the remains are dramatically damaged leaving very few interpretation clues to the visitor. In such cases a virtual presentation of the Cultural Heritage site can be of great help, specially for explaining the evolution of its status, giving sometimes sense to few spare stones. The definition of these digital representations deal with two crucial aspects: on the one hand the possibility of 3D surveying the relics in order to have an accurate geometrical image of the current status of the artifact; on the other hand the presence of historical sources both in form of written text or images, that once properly matched with the current geometrical data, may help to recreate digitally a set of 3D models representing visually the various historical phases (diachronic model, up to the current one. The core of this article is the definition of an integrated methodology that starts from an high-resolution digital survey of the remains of an ancient building and develops a coherent virtual reconstruction from different historical sources, suggesting a scalable method suitable to be re-used for generating a 4D (geometry + time model of the artifact. This approach has been experimented on the "Basilica di San Giovanni in Conca" in Milan, a very significant example for its complex historic evolution that combines evident historic values with an invisible presence inside the city.

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

    W. Schuhr; Lee, J. D.

    2015-01-01

    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 existin...

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

    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....

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

    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

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

    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 (R2 = 0.92) to the human oral adsorption than that of the Transwell culture (R2 = 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. (paper)

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

    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...

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

    Walther, Gerald

    2014-01-01

    In 2013, the first gun printed out of plastic by a 3D-printer was successfully fired in the US. 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 te...

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

    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. PMID:25520257

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

    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...... 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...... 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...

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

    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. PMID:25058941

  7. On-chip clearing of arrays of 3-D cell cultures and micro-tissues.

    Grist, S M; Nasseri, S S; Poon, T; Roskelley, C; Cheung, K C

    2016-07-01

    Three-dimensional (3-D) cell cultures are beneficial models for mimicking the complexities of in vivo tissues, especially in tumour studies where transport limitations can complicate response to cancer drugs. 3-D optical microscopy techniques are less involved than traditional embedding and sectioning, but are impeded by optical scattering properties of the tissues. Confocal and even two-photon microscopy limit sample imaging to approximately 100-200 μm depth, which is insufficient to image hypoxic spheroid cores. Optical clearing methods have permitted high-depth imaging of tissues without physical sectioning, but they are difficult to implement for smaller 3-D cultures due to sample loss in solution exchange. In this work, we demonstrate a microfluidic platform for high-throughput on-chip optical clearing of breast cancer spheroids using the SeeDB, Clear(T2), and ScaleSQ clearing methods. Although all three methods are able to effectively clear the spheroids, we find that SeeDB and ScaleSQ more effectively clear the sample than Clear(T2); however, SeeDB induces green autofluorescence while ScaleS causes sample expansion. Our unique on-chip implementation permits clearing arrays of 3-D cultures using perfusion while monitoring the 3-D cultures throughout the process, enabling visualization of the clearing endpoint as well as monitoring of transient changes that could induce image artefacts. Our microfluidic device is compatible with on-chip 3-D cell culture, permitting the use of on-chip clearing at the endpoint after monitoring the same spheroids during their culture. This on-chip method has the potential to improve readout from 3-D cultures, facilitating their use in cell-based assays for high-content drug screening and other applications. PMID:27493703

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

    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.

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

    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.

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

    Koeva, M. N.

    2016-01-01

    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 interact...

  11. 3D Modeling from Multi-views Images for Cultural Heritage in Wat-Pho, Thailand

    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.

  12. High Content Imaging (HCI on Miniaturized Three-Dimensional (3D Cell Cultures

    Pranav Joshi

    2015-12-01

    Full Text Available High content imaging (HCI is a multiplexed cell staining assay developed for better understanding of complex biological functions and mechanisms of drug action, and it has become an important tool for toxicity and efficacy screening of drug candidates. Conventional HCI assays have been carried out on two-dimensional (2D cell monolayer cultures, which in turn limit predictability of drug toxicity/efficacy in vivo; thus, there has been an urgent need to perform HCI assays on three-dimensional (3D cell cultures. Although 3D cell cultures better mimic in vivo microenvironments of human tissues and provide an in-depth understanding of the morphological and functional features of tissues, they are also limited by having relatively low throughput and thus are not amenable to high-throughput screening (HTS. One attempt of making 3D cell culture amenable for HTS is to utilize miniaturized cell culture platforms. This review aims to highlight miniaturized 3D cell culture platforms compatible with current HCI technology.

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

    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

  14. Hot embossing for fabrication of a microfluidic 3D cell culture platform.

    Jeon, Jessie S; Chung, Seok; Kamm, Roger D; Charest, Joseph L

    2011-04-01

    Clinically relevant studies of cell function in vitro require a physiologically-representative microenvironment possessing aspects such as a 3D extracellular matrix (ECM) and controlled biochemical and biophysical parameters. A polydimethylsiloxane (PDMS) microfluidic system with a 3D collagen gel has previously served for analysis of factors inducing different responses of cells in a 3D microenvironment under controlled biochemical and biophysical parameters. In the present study, applying the known commercially-viable manufacturing methods to a cyclic olefin copolymer (COC) material resulted in a microfluidic device with enhanced 3D gel capabilities, controlled surface properties, and improved potential to serve high-volume applications. Hot embossing and roller lamination molded and sealed the microfluidic device. A combination of oxygen plasma and thermal treatments enhanced the sealing, ensured proper placement of the 3D gel, and created controlled and stable surface properties within the device. Culture of cells in the new device indicated no adverse effects of the COC material or processing as compared to previous PDMS devices. The results demonstrate a methodology to transition microfluidic devices for 3D cell culture from scientific research to high-volume applications with broad clinical impact. PMID:21113663

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

    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.

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

    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.

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

    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. PMID:26978075

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

    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. PMID:26978075

  19. Self-assembling Fmoc dipeptide hydrogel for in situ 3D cell culturing

    Akpe Victor; Rydholm Susanna; Liebmann Thomas; Brismar Hjalmar

    2007-01-01

    Abstract Background Conventional cell culture studies have been performed on 2D surfaces, resulting in flat, extended cell growth. More relevant studies are desired to better mimic 3D in vivo tissue growth. Such realistic environments should be the aim of any cell growth study, requiring new methods for culturing cells in vitro. Cell biology is also tending toward miniaturization for increased efficiency and specificity. This paper discusses the application of a self-assembling peptide-derive...

  20. 3D Hepatic Cultures Simultaneously Maintain Primary Hepatocyte and Liver Sinusoidal Endothelial Cell Phenotypes

    Kim, Yeonhee; Rajagopalan, Padmavathy

    2010-01-01

    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, detoxification and

  1. 3D hepatic cultures simultaneously maintain primary hepatocyte and liver sinusoidal endothelial cell phenotypes.

    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

  2. Self-assembling Fmoc dipeptide hydrogel for in situ 3D cell culturing

    Liebmann, Thomas; Rydholm, Susanna; Akpe, Victor; Brismar, Hjalmar

    2007-01-01

    Background Conventional cell culture studies have been performed on 2D surfaces, resulting in flat, extended cell growth. More relevant studies are desired to better mimic 3D in vivo tissue growth. Such realistic environments should be the aim of any cell growth study, requiring new methods for culturing cells in vitro. Cell biology is also tending toward miniaturization for increased efficiency and specificity. This paper discusses the application of a self-assembling peptide-derived hydrogel for use as a 3D cell culture scaffold at the microscale. Results Phenylalanine derivative hydrogel formation was seen to occur in multiple dispersion media. Cells were immobilized in situ within microchambers designed for cell analysis. Use of the highly biocompatible hydrogel components and simplistic procedures significantly reduced the cytotoxic effects seen with alternate 3D culture materials and microstructure loading methods. Cells were easily immobilized, sustained and removed from microchambers. Differences in growth morphology were seen in the cultured cells, owing to the 3-dimentional character of the gel structure. Degradation improved the removal of hydrogel from the microstructures, permitting reuse of the analysis platforms. Conclusion Self-assembling diphenylalanine derivative hydrogel provided a method to dramatically reduce the typical difficulties of microculture formation. Effective generation of patterned 3D cultures will lead to improved cell study results by better modeling in vivo growth environments and increasing efficiency and specificity of cell studies. Use of simplified growth scaffolds such as peptide-derived hydrogel should be seen as highly advantageous and will likely become more commonplace in cell culture methodology. PMID:18070345

  3. Self-assembling Fmoc dipeptide hydrogel for in situ 3D cell culturing

    Akpe Victor

    2007-12-01

    Full Text Available Abstract Background Conventional cell culture studies have been performed on 2D surfaces, resulting in flat, extended cell growth. More relevant studies are desired to better mimic 3D in vivo tissue growth. Such realistic environments should be the aim of any cell growth study, requiring new methods for culturing cells in vitro. Cell biology is also tending toward miniaturization for increased efficiency and specificity. This paper discusses the application of a self-assembling peptide-derived hydrogel for use as a 3D cell culture scaffold at the microscale. Results Phenylalanine derivative hydrogel formation was seen to occur in multiple dispersion media. Cells were immobilized in situ within microchambers designed for cell analysis. Use of the highly biocompatible hydrogel components and simplistic procedures significantly reduced the cytotoxic effects seen with alternate 3D culture materials and microstructure loading methods. Cells were easily immobilized, sustained and removed from microchambers. Differences in growth morphology were seen in the cultured cells, owing to the 3-dimentional character of the gel structure. Degradation improved the removal of hydrogel from the microstructures, permitting reuse of the analysis platforms. Conclusion Self-assembling diphenylalanine derivative hydrogel provided a method to dramatically reduce the typical difficulties of microculture formation. Effective generation of patterned 3D cultures will lead to improved cell study results by better modeling in vivo growth environments and increasing efficiency and specificity of cell studies. Use of simplified growth scaffolds such as peptide-derived hydrogel should be seen as highly advantageous and will likely become more commonplace in cell culture methodology.

  4. 3D Hydrogel Scaffolds for Articular Chondrocyte Culture and Cartilage Generation.

    Smeriglio, Piera; Lai, Janice H; Yang, Fan; Bhutani, Nidhi

    2015-01-01

    Human articular cartilage is highly susceptible to damage and has limited self-repair and regeneration potential. Cell-based strategies to engineer cartilage tissue offer a promising solution to repair articular cartilage. To select the optimal cell source for tissue repair, it is important to develop an appropriate culture platform to systematically examine the biological and biomechanical differences in the tissue-engineered cartilage by different cell sources. Here we applied a three-dimensional (3D) biomimetic hydrogel culture platform to systematically examine cartilage regeneration potential of juvenile, adult, and osteoarthritic (OA) chondrocytes. The 3D biomimetic hydrogel consisted of synthetic component poly(ethylene glycol) and bioactive component chondroitin sulfate, which provides a physiologically relevant microenvironment for in vitro culture of chondrocytes. In addition, the scaffold may be potentially used for cell delivery for cartilage repair in vivo. Cartilage tissue engineered in the scaffold can be evaluated using quantitative gene expression, immunofluorescence staining, biochemical assays, and mechanical testing. Utilizing these outcomes, we were able to characterize the differential regenerative potential of chondrocytes of varying age, both at the gene expression level and in the biochemical and biomechanical properties of the engineered cartilage tissue. The 3D culture model could be applied to investigate the molecular and functional differences among chondrocytes and progenitor cells from different stages of normal or aberrant development. PMID:26484414

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

    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.

  6. Human disc cells in monolayer vs 3D culture: cell shape, division and matrix formation

    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.

  7. Human dental pulp stem cells produce mineralized matrix in 2D and 3D cultures

    M. Riccio; Resca, E.; Maraldi, T; Pisciotta, A.; Ferrari, A; Bruzzesi, G.; De Pol, A.

    2010-01-01

    The aim of this study was to characterize the in vitro osteogenic differentiation of dental pulp stem cells (DPSCs) in 2D cultures and 3D biomaterials. DPSCs, separated from dental pulp by enzymatic digestion, and isolated by magnetic cell sorting were differentiated toward osteogenic lineage on 2D surface by using an osteogenic medium. During the differentiation process, DPSCs express specific bone proteins like Runx-2, Osx, OPN and OCN with a sequential expression, analogous to those occurr...

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

    Sonntag, F.; Schmieder, F.; Ströbel, J.; Grünzner, S.; Busek, M.; Günther, K.; Steege, T.; Polk, C.; Klotzbach, U.

    2016-03-01

    The miniaturization, rapid prototyping and automation of lab-on-a-chip technology play nowadays a very important role. Lab-on-a-chip technology is successfully implemented not only for environmental analysis and medical diagnostics, but also as replacement of animals used for the testing of substances in the pharmaceutical and cosmetics industries. For that purpose the Fraunhofer IWS and partners developed a lab-on-a-chip platform for perfused cell-based assays in the last years, which includes different micropumps, valves, channels, reservoirs and customized cell culture modules. This technology is already implemented for the characterization of different human cell cultures and organoids, like skin, liver, endothelium, hair follicle and nephron. The advanced universal lab-on-a-chip platform for complex, perfused 3D cell cultures is divided into a multilayer basic chip with integrated micropump and application-specific 3D printed cell culture modules. Moreover a technology for surface modification of the printed cell culture modules by laser micro structuring and a complex and flexibly programmable controlling device based on an embedded Linux system was developed. A universal lab-on-a-chip platform with an optional oxygenator and a cell culture module for cubic scaffolds as well as first cell culture experiments within the cell culture device will be presented. The module is designed for direct interaction with robotic dispenser systems. This offers the opportunity to combine direct organ printing of cells and scaffolds with the microfluidic cell culture module. The characterization of the developed system was done by means of Micro-Particle Image Velocimetry (μPIV) and an optical oxygen measuring system.

  9. MAPLE deposition of 3D micropatterned polymeric substrates for cell culture

    Paun, Irina Alexandra, E-mail: irina.paun@physics.pub.ro [National Institute for Laser, Plasma and Radiation Physics, RO-077125, Magurele, Bucharest (Romania); Faculty of Applied Sciences, University Politehnica of Bucharest, RO-060042, Bucharest (Romania); Mihailescu, Mona [Faculty of Applied Sciences, University Politehnica of Bucharest, RO-060042, Bucharest (Romania); Calenic, Bogdan [Department of Biochemistry, Faculty of Dentistry, UMF Carol Davila, Bucharest (Romania); Luculescu, Catalin Romeo [National Institute for Laser, Plasma and Radiation Physics, RO-077125, Magurele, Bucharest (Romania); Greabu, Maria [Department of Biochemistry, Faculty of Dentistry, UMF Carol Davila, Bucharest (Romania); Dinescu, Maria, E-mail: dinescum@nipne.ro [National Institute for Laser, Plasma and Radiation Physics, RO-077125, Magurele, Bucharest (Romania)

    2013-08-01

    3D micropatterned poly(lactide-co-glycolide)/polyurethane (PLGA/PU) substrates were produced by MAPLE deposition through masks and used for regulating the behavior of oral keratinocyte stem cells in response to topography. Flat PLGA/PU substrates were produced for comparison. 3D imaging of the PLGA/PU substrates and of the cultured cells was performed by Digital Holographic Microscopy. The micropatterns were in the shape of squares of 50 × 50 and 80 × 80 μm{sup 2} areas, ∼1.8 μm in height and separated by 20 μm wide channels. It was found that substrate topography guided the adhesion of the cultured cells: on the smooth substrates the cells adhered randomly and showed no preferred orientation; in contrast, on the micropatterned substrates the cells adhered preferentially onto the squares and not in the separating channels. Furthermore, key properties of the cells (size, viability, proliferation rate and stem cell marker expression) did not show any dependence on substrate topography. The size of the cultured cells, their viability, the proportions of actively/slow proliferating cells, as well as the stem cell markers expressions, were similar for both flat and micropatterned substrates. Finally, it was found that the cells cultured on the PLGA/PU substrates deposited by MAPLE exhibited similar properties as the controls (i.e. cells cultured on glass slides), indicating the capability of the former to preserve the properties of the keratinocyte stem cells.

  10. MAPLE deposition of 3D micropatterned polymeric substrates for cell culture

    3D micropatterned poly(lactide-co-glycolide)/polyurethane (PLGA/PU) substrates were produced by MAPLE deposition through masks and used for regulating the behavior of oral keratinocyte stem cells in response to topography. Flat PLGA/PU substrates were produced for comparison. 3D imaging of the PLGA/PU substrates and of the cultured cells was performed by Digital Holographic Microscopy. The micropatterns were in the shape of squares of 50 × 50 and 80 × 80 μm2 areas, ∼1.8 μm in height and separated by 20 μm wide channels. It was found that substrate topography guided the adhesion of the cultured cells: on the smooth substrates the cells adhered randomly and showed no preferred orientation; in contrast, on the micropatterned substrates the cells adhered preferentially onto the squares and not in the separating channels. Furthermore, key properties of the cells (size, viability, proliferation rate and stem cell marker expression) did not show any dependence on substrate topography. The size of the cultured cells, their viability, the proportions of actively/slow proliferating cells, as well as the stem cell markers expressions, were similar for both flat and micropatterned substrates. Finally, it was found that the cells cultured on the PLGA/PU substrates deposited by MAPLE exhibited similar properties as the controls (i.e. cells cultured on glass slides), indicating the capability of the former to preserve the properties of the keratinocyte stem cells.

  11. The potential of 3D techniques for cultural heritage object documentation

    Bitelli, Gabriele; Girelli, Valentina A.; Remondino, Fabio; Vittuari, Luca

    2007-01-01

    The generation of 3D models of objects has become an important research point in many fields of application like industrial inspection, robotics, navigation and body scanning. Recently the techniques for generating photo-textured 3D digital models have interested also the field of Cultural Heritage, due to their capability to combine high precision metrical information with a qualitative and photographic description of the objects. In fact this kind of product is a fundamental support for documentation, studying and restoration of works of art, until a production of replicas by fast prototyping techniques. Close-range photogrammetric techniques are nowadays more and more frequently used for the generation of precise 3D models. With the advent of automated procedures and fully digital products in the 1990s, it has become easier to use and cheaper, and nowadays a wide range of commercial software is available to calibrate, orient and reconstruct objects from images. This paper presents the complete process for the derivation of a photorealistic 3D model of an important basalt stela (about 70 x 60 x 25 cm) discovered in the archaeological site of Tilmen Höyük, in Turkey, dating back to 2nd mill. BC. We will report the modeling performed using passive and active sensors and the comparison of the achieved results.

  12. Human dental pulp stem cells produce mineralized matrix in 2D and 3D cultures

    M. Riccio

    2010-11-01

    Full Text Available The aim of this study was to characterize the in vitro osteogenic differentiation of dental pulp stem cells (DPSCs in 2D cultures and 3D biomaterials. DPSCs, separated from dental pulp by enzymatic digestion, and isolated by magnetic cell sorting were differentiated toward osteogenic lineage on 2D surface by using an osteogenic medium. During the differentiation process, DPSCs express specific bone proteins like Runx-2, Osx, OPN and OCN with a sequential expression, analogous to those occurring during osteoblast differentiation, and produce extracellular calcium deposits. In order to differentiate cells in a 3D space that mimes the physiological environment, DPSCs were cultured in two distinct bioscaffolds, MatrigelTM and Collagen sponge. With the addition of a third dimension, osteogenic differentiation and mineralized extracellular matrix production significantly improved. In particular, in MatrigelTM DPSCs differentiated with osteoblast/osteocyte characteristics and connected by gap junction, and therefore formed calcified nodules with a 3D intercellular network. Furthermore, DPSCs differentiated in collagen sponge actively secrete human type I collagen micro-fibrils and form calcified matrix containing trabecular-like structures. These neo-formed DPSCs-scaffold devices may be used in regenerative surgical applications in order to resolve pathologies and traumas characterized by critical size bone defects.

  13. Disulfide-Based Diblock Copolymer Worm Gels: A Wholly-Synthetic Thermoreversible 3D Matrix for Sheet-Based Cultures

    Simon, Karen Alambra; Warren, Nicholas J.; Mosadegh, Bobak; Mohammady, Marym R.; Whitesides, George McClelland; Armes, Steven P.

    2015-01-01

    It is well-known that 3D in vitro cell cultures provide a much better model than 2D cell cultures for understanding the in vivo microenvironment of cells. However, significant technical challenges in handling and analyzing 3D cell cultures remain, which currently limits their widespread application. Herein, we demonstrate the application of wholly synthetic thermoresponsive block copolymer worms in sheet-based 3D cell culture. These worms form a soft, free-standing gel reversibly at 20–37 °C,...

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

    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.

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

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

    2014-01-01

    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. PMID:24670718

  16. Towards a 3d Based Platform for Cultural Heritage Site Survey and Virtual Exploration

    Seinturier, J.; Riedinger, C.; Mahiddine, A.; Peloso, D.; Boï, J.-M.; Merad, D.; Drap, P.

    2013-07-01

    This paper present a 3D platform that enables to make both cultural heritage site survey and its virtual exploration. It provides a single and easy way to use framework for merging multi scaled 3D measurements based on photogrammetry, documentation produced by experts and the knowledge of involved domains leaving the experts able to extract and choose the relevant information to produce the final survey. Taking into account the interpretation of the real world during the process of archaeological surveys is in fact the main goal of a survey. New advances in photogrammetry and the capability to produce dense 3D point clouds do not solve the problem of surveys. New opportunities for 3D representation are now available and we must to use them and find new ways to link geometry and knowledge. The new platform is able to efficiently manage and process large 3D data (points set, meshes) thanks to the implementation of space partition methods coming from the state of the art such as octrees and kd-trees and thus can interact with dense point clouds (thousands to millions of points) in real time. The semantisation of raw 3D data relies on geometric algorithms such as geodetic path computation, surface extraction from dense points cloud and geometrical primitive optimization. The platform provide an interface that enables expert to describe geometric representations of interesting objects like ashlar blocs, stratigraphic units or generic items (contour, lines, … ) directly onto the 3D representation of the site and without explicit links to underlying algorithms. The platform provide two ways for describing geometric representation. If oriented photographs are available, the expert can draw geometry on a photograph and the system computes its 3D representation by projection on the underlying mesh or the points cloud. If photographs are not available or if the expert wants to only use the 3D representation then he can simply draw objects shape on it. When 3D

  17. 3D culture of ovarian follicles: a system towards their engineering?

    Zuccotti, Maurizio; Merico, Valeria; Rebuzzini, Paola; Belli, Martina; Vigone, Giulia; Mulas, Francesca; Fassina, Lorenzo; Wruck, Wasco; Adjaye, James; Bellazzi, Riccardo; Garagna, Silvia

    2015-01-01

    Infertility in women is a health priority. Designing a robust culture protocol capable of attaining complete follicle growth is an exciting challenge, for its potential clinical applications, but also as a model to observe and closely study the sequence of molecular events that lie behind the intricate relationship existing between the oocyte and surrounding follicle cells. Here, we describe the procedures used to maintain the ovarian follicle 3D architecture employing a variety of in vitro systems and several types of matrices. Collagen and alginate are the matrices that led to better results, including proof-of-concept of full-term development. Pioneer in its kind, these studies underlie the drawbacks encountered and the need for a culture system that allows more quantitative analyses and predictions, projecting the culture of the ovarian follicle into the realm of tissue engineering. PMID:26505254

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

    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.

  19. 3D cell culture systems modeling tumor growth determinants in cancer target discovery.

    Thoma, Claudio R; Zimmermann, Miriam; Agarkova, Irina; Kelm, Jens M; Krek, Wilhelm

    2014-04-01

    Phenotypic heterogeneity of cancer cells, cell biological context, heterotypic crosstalk and the microenvironment are key determinants of the multistep process of tumor development. They sign responsible, to a significant extent, for the limited response and resistance of cancer cells to molecular-targeted therapies. Better functional knowledge of the complex intra- and intercellular signaling circuits underlying communication between the different cell types populating a tumor tissue and of the systemic and local factors that shape the tumor microenvironment is therefore imperative. Sophisticated 3D multicellular tumor spheroid (MCTS) systems provide an emerging tool to model the phenotypic and cellular heterogeneity as well as microenvironmental aspects of in vivo tumor growth. In this review we discuss the cellular, chemical and physical factors contributing to zonation and cellular crosstalk within tumor masses. On this basis, we further describe 3D cell culture technologies for growth of MCTS as advanced tools for exploring molecular tumor growth determinants and facilitating drug discovery efforts. We conclude with a synopsis on technological aspects for on-line analysis and post-processing of 3D MCTS models. PMID:24636868

  20. Micro 3D cell culture systems for cellular behavior studies: Culture matrices, devices, substrates, and in-situ sensing methods.

    Choi, Jonghoon; Lee, Eun Kyu; Choo, Jaebum; Yuh, Junhan; Hong, Jong Wook

    2015-09-01

    Microfabricated systems equipped with 3D cell culture devices and in-situ cellular biosensing tools can be a powerful bionanotechnology platform to investigate a variety of biomedical applications. Various construction substrates such as plastics, glass, and paper are used for microstructures. When selecting a construction substrate, a key consideration is a porous microenvironment that allows for spheroid growth and mimics the extracellular matrix (ECM) of cell aggregates. Various bio-functionalized hydrogels are ideal candidates that mimic the natural ECM for 3D cell culture. When selecting an optimal and appropriate microfabrication method, both the intended use of the system and the characteristics and restrictions of the target cells should be carefully considered. For highly sensitive and near-cell surface detection of excreted cellular compounds, SERS-based microsystems capable of dual modal imaging have the potential to be powerful tools; however, the development of optical reporters and nanoprobes remains a key challenge. We expect that the microsystems capable of both 3D cell culture and cellular response monitoring would serve as excellent tools to provide fundamental cellular behavior information for various biomedical applications such as metastasis, wound healing, high throughput screening, tissue engineering, regenerative medicine, and drug discovery and development. PMID:26358782

  1. The Cultural Divide: Exponential Growth in Classical 2D and Metabolic Equilibrium in 3D Environments

    Wrzesinski, Krzysztof; Rogowska-Wrzesinska, Adelina; Kanlaya, Rattiyaporn;

    2014-01-01

    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...... 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...

  2. Efficient Use of Video for 3d Modelling of Cultural Heritage Objects

    Alsadik, B.; Gerke, M.; Vosselman, G.

    2015-03-01

    Currently, there is a rapid development in the techniques of the automated image based modelling (IBM), especially in advanced structure-from-motion (SFM) and dense image matching methods, and camera technology. One possibility is to use video imaging to create 3D reality based models of cultural heritage architectures and monuments. Practically, video imaging is much easier to apply when compared to still image shooting in IBM techniques because the latter needs a thorough planning and proficiency. However, one is faced with mainly three problems when video image sequences are used for highly detailed modelling and dimensional survey of cultural heritage objects. These problems are: the low resolution of video images, the need to process a large number of short baseline video images and blur effects due to camera shake on a significant number of images. In this research, the feasibility of using video images for efficient 3D modelling is investigated. A method is developed to find the minimal significant number of video images in terms of object coverage and blur effect. This reduction in video images is convenient to decrease the processing time and to create a reliable textured 3D model compared with models produced by still imaging. Two experiments for modelling a building and a monument are tested using a video image resolution of 1920×1080 pixels. Internal and external validations of the produced models are applied to find out the final predicted accuracy and the model level of details. Related to the object complexity and video imaging resolution, the tests show an achievable average accuracy between 1 - 5 cm when using video imaging, which is suitable for visualization, virtual museums and low detailed documentation.

  3. Low-level laser therapy in 3D cell culture model using gingival fibroblasts.

    Basso, Fernanda G; Soares, Diana G; de Souza Costa, Carlos Alberto; Hebling, Josimeri

    2016-07-01

    Besides extensive data about the effects of low-level laser therapy (LLLT) on different cell types, so far, these results were obtained from monolayer cell culture models, which have limitations in terms of cell morphology and phenotype expression. Therefore, for better in vitro evaluation of the effects of LLLT, this study was performed with a 3D cell culture model, where gingival fibroblasts were seeded in collagen matrix. Cells isolated from a healthy patient were seeded in wells of 24-well plates with culture medium (DMEM) supplemented with 10 % fetal bovine serum and collagen type I solution. After 5 days, a serum-free DMEM was added to the matrices with cells that were subjected or not to three consecutive irradiations of LLLT by means of the LaserTABLE diode device (780 nm, 25 mW) at 0.5, 1.5, and 3 J/cm(2). Twenty-four hours after the last irradiation, cell viability and morphology as well as gene expression of growth factors were assessed. Histological evaluation of matrices demonstrated uniform distribution and morphology of gingival fibroblasts within the collagen matrix. LLLT at 3 J/cm(2) increased gingival fibroblast viability. Enhanced gene expression of hCOL-I and hEGF was observed for 0.5 J/cm(2), while no significant changes were detected for the other irradiation densities tested. In conclusion, LLLT promoted biostimulation of gingival fibroblasts seeded in a 3D cell culture model, demonstrating that this model can be applied for phototherapy studies and that LLLT could penetrate the collagen matrix to increase cell functions related to tissue repair. PMID:27126408

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

    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.

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

    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.

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

    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

  7. Endothelial cells stimulate growth of normal and cancerous breast epithelial cells in 3D culture

    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.

    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. PMID:27552143

  9. Drug penetration and metabolism in 3D cell cultures treated in a 3D printed fluidic device: assessment of irinotecan via MALDI imaging mass spectrometry.

    LaBonia, Gabriel J; Lockwood, Sarah Y; Heller, Andrew A; Spence, Dana M; Hummon, Amanda B

    2016-06-01

    Realistic in vitro models are critical in the drug development process. In this study, a novel in vitro platform is employed to assess drug penetration and metabolism. This platform, which utilizes a 3D printed fluidic device, allows for dynamic dosing of three dimensional cell cultures, also known as spheroids. The penetration of the chemotherapeutic irinotecan into HCT 116 colon cancer spheroids was examined with MALDI imaging mass spectrometry (IMS). The active metabolite of irinotecan, SN-38, was also detected. After twenty-four hours of treatment, SN-38 was concentrated to the outside of the spheroid, a region of actively dividing cells. The irinotecan prodrug localization contrasted with SN-38 and was concentrated to the necrotic core of the spheroids, a region containing mostly dead and dying cells. These results demonstrate that this unique in vitro platform is an effective means to assess drug penetration and metabolism in 3D cell cultures. This innovative system can have a transformative impact on the preclinical evaluation of drug candidates due to its cost effectiveness and high throughput. PMID:27198560

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

    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.

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

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

    2016-01-01

    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. PMID:27215466

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

    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.

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

    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.

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

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

    2016-01-01

    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. PMID:27215466

  15. AlgiMatrix™ Based 3D Cell Culture System as an In-Vitro Tumor Model for Anticancer Studies

    Godugu, Chandraiah; Patel, Apurva R.; Desai, Utkarsh; Andey, Terrick; Sams, Alexandria; Singh, Mandip

    2013-01-01

    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 co...

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

    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.

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

    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 E2 (PGE2) 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.

  18. A Multifunctional 3D Co-Culture System for Studies of Mammary Tissue Morphogenesis and Stem Cell Biology

    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 sc...

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

    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

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

    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.

  1. Dual targeting of EGFR and focal adhesion kinase in 3D grown HNSCC cell cultures

    Purpose: Epidermal growth factor receptor (EGFR) and focal adhesion kinase (FAK) show frequent overexpression and hyperactivity in various human malignancies including head and neck squamous cell carcinomas (HNSCC). To examine effects of dual EGFR/FAK inhibition on cellular radiosensitivity of HNSCC cells in a more physiological environment, we employed a previously established laminin-rich extracellular matrix (lrECM) based three-dimensional (3D) cell culture model. Materials and methods: UTSCC15 and SAS HNSCC cell lines stably transfected with EGFR-CFP or CFP were used. Single or combined EGFR (Cetuximab, siRNA) and FAK (TAE226, siRNA) inhibition were accomplished prior to measuring clonogenic survival and protein expression and phosphorylation. Immunofluorescence enabled visualization of EGFR-CFP and FAK. Results: Cetuximab resulted in higher radiosensitization in EGFR-CFP overexpressing cell lines than CFP controls. Single EGFR or FAK inhibition mediated radiosensitization, while dual EGFR/FAK targeting further augmented this effect. Despite signaling alterations upon Cetuximab and siRNA knockdown, analysis of protein expression and phosphorylation indicates EGFR and FAK signaling coexistence without obvious overlap. Conclusions: Combined EGFR/FAK targeting yielded stronger radiosensitization than either approach alone, which might be based on non-overlapping downstream signaling. Whether dual targeting of EGFR and FAK can reasonably be combined with radiotherapy and chemotherapy needs clarification.

  2. Accuracy of cultural heritage 3D models by RPAS and terrestrial photogrammetry

    Bolognesi, M.; Furini, A.; Russo, V.; Pellegrinelli, A.; Russo, P.

    2014-06-01

    The combined use of high-resolution digital images taken from ground as well as from RPAS (Remotely Piloted Aircraft Systems) have significantly increased the potential of close range digital photogrammetry applications in Cultural Heritage surveying and modeling. It is in fact possible, thanks to SfM (Structure from Motion), to simultaneously process great numbers of aerial and terrestrial images for the production of a dense point cloud of an object. In order to analyze the accuracy of results, we started numerous tests based on the comparison between 3D digital models of a monumental complex realized by the integration of aerial and terrestrial photogrammetry and an accurate TLS (Terrestrial Laser Scanner) reference model of the same object. A lot of digital images of a renaissance castle, assumed as test site, have been taken both by ground level and by RPAS at different distances and flight altitudes and with different flight patterns. As first step of the experimentation, the images were previously processed with Agisoft PhotoScan, one of the most popular photogrammetric software. The comparison between the photogrammetric DSM of the monument and a TLS reference one was carried out by evaluating the average deviation between the points belonging to the two entities, both globally and locally, on individual façades and architectural elements (sections and particular). In this paper the results of the first test are presented. A good agreement between photogrammetric and TLS digital models of the castle is pointed out.

  3. Hot embossing for fabrication of a microfluidic 3D cell culture platform

    Jeon, Jessie S.; Chung, Seok; Kamm, Roger D.; Charest, Joseph L.

    2011-01-01

    Clinically relevant studies of cell function in vitro require a physiologically-representative microenvironment possessing aspects such as a 3D extracellular matrix (ECM) and controlled biochemical and biophysical parameters. A polydimethylsiloxane (PDMS) microfluidic system with a 3D collagen gel has previously served for analysis of factors inducing different responses of cells in a 3D microenvironment under controlled biochemical and biophysical parameters. In the present study, applying t...

  4. Hot embossing for fabrication of a microfluidic 3D cell culture

    Jeon, Jessie S.; Chung, Seok; Kamm, Roger Dale; Charest, Joseph L.

    2010-01-01

    Clinically relevant studies of cell function in vitro require a physiologically-representative microenvironment possessing aspects such as a 3D extracellular matrix (ECM) and controlled biochemical and biophysical parameters. A polydimethylsiloxane (PDMS) microfluidic system with a 3D collagen gel has previously served for analysis of factors inducing different responses of cells in a 3D microenvironment under controlled biochemical and biophysical parameters. In the present study, applying t...

  5. Disulfide-Based Diblock Copolymer Worm Gels: A Wholly-Synthetic Thermoreversible 3D Matrix for Sheet-Based Cultures.

    Simon, Karen A; Warren, Nicholas J; Mosadegh, Bobak; Mohammady, Marym R; Whitesides, George M; Armes, Steven P

    2015-12-14

    It is well-known that 3D in vitro cell cultures provide a much better model than 2D cell cultures for understanding the in vivo microenvironment of cells. However, significant technical challenges in handling and analyzing 3D cell cultures remain, which currently limits their widespread application. Herein, we demonstrate the application of wholly synthetic thermoresponsive block copolymer worms in sheet-based 3D cell culture. These worms form a soft, free-standing gel reversibly at 20-37 °C, which can be rapidly converted into a free-flowing dispersion of spheres on cooling to 5 °C. Functionalization of the worms with disulfide groups was found to be essential for ensuring sufficient mechanical stability of these hydrogels to enable long-term cell culture. These disulfide groups are conveniently introduced via statistical copolymerization of a disulfide-based dimethacrylate under conditions that favor intramolecular cyclization and subsequent thiol/disulfide exchange leads to the formation of reversible covalent bonds between adjacent worms within the gel. This new approach enables cells to be embedded within micrometer-thick slabs of gel with good viability, permits cell culture for at least 12 days, and facilitates recovery of viable cells from the gel simply by incubating the culture in buffer at 4 °C (thus, avoiding the enzymatic degradation required for cell harvesting when using commercial protein-based gels, such as Matrigel). PMID:26509930

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

    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...

  7. Epithelial-to-mesenchymal transition in pancreatic ductal adenocarcinoma: Characterization in a 3D-cell culture model

    Gagliano, Nicoletta; Celesti, Giuseppe; Tacchini, Lorenza; Pluchino, Stefano; Sforza, Chiarella; Rasile, Marco; Valerio, Vincenza; Laghi, Luigi; Conte, Vincenzo; Procacci, Patrizia

    2016-01-01

    AIM: To analyze the effect of three-dimensional (3D)-arrangement on the expression of epithelial-to-mesenchymal transition markers in pancreatic adenocarcinoma (PDAC) cells. METHODS: HPAF-II, HPAC, and PL45 PDAC cells were cultured in either 2D-monolayers or 3D-spheroids. Ultrastructure was analyzed by transmission electron microscopy. The expression of E-cadherin, β-catenin, N-cadherin, collagen type I (COL-I), vimentin, α-smooth muscle actin (αSMA), and podoplanin was assayed by confocal microscopy in cells cultured on 12-mm diameter round coverslips and in 3D-spheroids. Gene expression for E-cadherin, Snail, Slug, Twist, Zeb1, and Zeb2 was quantified by real-time PCR. E-cadherin protein level and its electrophoretic pattern were studied by Western blot in cell lysates obtained from cells grown in 2D-monolayers and 3D-spheroids. RESULTS: The E-cadherin/β-catenin complex was expressed in a similar way in plasma membrane cell boundaries in both 2D-monolayers and 3D-spheroids. E-cadherin increased in lysates obtained from 3D-spheroids, while cleavage fragments were more evident in 2D-monolayers. N-cadherin expression was observed in very few PDAC cells grown in 2D-monolayers, but was more evident in 3D-spheroids. Some cells expressing COL-I were observed in 3D-spheroids. Podoplanin, expressed in collectively migrating cells, and αSMA were similarly expressed in both experimental conditions. The concomitant maintenance of the E-cadherin/β-catenin complex at cell boundaries supports the hypothesis of a collective migration for these cells, which is consistent with podoplanin expression. CONCLUSION: We show that a 3D-cell culture model could provide deeper insight into understanding the biology of PDAC and allow for the detection of marked differences in the phenotype of PDAC cells grown in 3D-spheroids. PMID:27182158

  8. Surface Acoustic Waves (SAW-Based Biosensing for Quantification of Cell Growth in 2D and 3D Cultures

    Tao Wang

    2015-12-01

    Full Text Available Detection and quantification of cell viability and growth in two-dimensional (2D and three-dimensional (3D cell cultures commonly involve harvesting of cells and therefore requires a parallel set-up of several replicates for time-lapse or dose–response studies. Thus, developing a non-invasive and touch-free detection of cell growth in longitudinal studies of 3D tumor spheroid cultures or of stem cell regeneration remains a major unmet need. Since surface acoustic waves (SAWs permit mass loading-based biosensing and have been touted due to their many advantages including low cost, small size and ease of assembly, we examined the potential of SAW-biosensing to detect and quantify cell growth. Herein, we demonstrate that a shear horizontal-surface acoustic waves (SH-SAW device comprising two pairs of resonators consisting of interdigital transducers and reflecting fingers can be used to quantify mass loading by the cells in suspension as well as within a 3D cell culture platform. A 3D COMSOL model was built to simulate the mass loading response of increasing concentrations of cells in suspension in the polydimethylsiloxane (PDMS well in order to predict the characteristics and optimize the design of the SH-SAW biosensor. The simulated relative frequency shift from the two oscillatory circuit systems (one of which functions as control were found to be concordant to experimental data generated with RAW264.7 macrophage and A549 cancer cells. In addition, results showed that SAW measurements per se did not affect viability of cells. Further, SH-SAW biosensing was applied to A549 cells cultured on a 3D electrospun nanofiber scaffold that generate tumor spheroids (tumoroids and the results showed the device's ability to detect changes in tumor spheroid growth over the course of eight days. Taken together, these results demonstrate the use of SH-SAW device for detection and quantification of cell growth changes over time in 2D suspension cultures and in

  9. 5D Modelling: An Efficient Approach for Creating Spatiotemporal Predictive 3D Maps of Large-Scale Cultural Resources

    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

  10. Infrared imaging of MDA-MB-231 breast cancer cell line phenotypes in 2D and 3D cultures.

    Smolina, Margarita; Goormaghtigh, Erik

    2015-04-01

    One current challenge in the field of breast cancer infrared imaging is the identification of carcinoma cell subtypes in the tissue. Neither sequencing nor immunochemistry is currently able to provide a cell by cell thorough classification. The latter is needed to build accurate statistical models capable of recognizing the diversity of breast cancer cell lines that may be present in a tissue section. One possible approach for overcoming this problem is to obtain the IR spectral signature of well-characterized tumor cell lines in culture. Cultures in three-dimensional matrices appear to generate an environment that mimics better the in vivo environment. There are, at present, series of breast cancer cell lines that have been thoroughly characterized in two- and three-dimensional (2D and 3D) cultures by full transcriptomics analyses. In this work, we describe the methods used to grow, to process, and to characterize a triple-negative breast cancer cell line, MDA-MB-231, in 3D laminin-rich extracellular matrix (lrECM) culture and compare it with traditional monolayer cultures and tissue sections. While unsupervised analyses did not completely separate spectra of cells grown in 2D from 3D lrECM cultures, a supervised statistical analysis resulted in an almost perfect separation. When IR spectral responses of epithelial tumor cells from clinical triple-negative breast carcinoma samples were added to these data, a principal component analysis indicated that they cluster closer to the spectra of 3D culture cells than to the spectra of cells grown on a flat plastic substrata. This result is encouraging because of correlating well-characterized cell line features with clinical biopsies. PMID:25568895

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

    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.

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

    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

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

    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.

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

    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.

  15. The relevance of using 3D cell cultures, in addition to 2D monolayer cultures, when evaluating breast cancer drug sensitivity and resistance.

    O'Driscoll, Lorraine

    2016-01-01

    PUBLISHED 2016 Jun 10. doi: 10.18632/oncotarget.9935. [Epub ahead of print] Solid tumours naturally grow in 3D wherein the spatial arrangement of cells affects how they interact with each other. This suggests that 3D cell culture may mimic the natural in vivo setting better than traditional monolayer (2D) cell culture, where cells are grown attached to plastic. Here, using HER2-positive breast cancer cell lines as models (BT474, HCC1954, EFM192A), the effects of culturing c...

  16. 3D Visualization of Cultural Heritage Artefacts with Virtual Reality devices

    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.

  17. 3D Realisation of Cultural Heritage UNESCO world heritage site in Saltaire, Bradford

    Unver, Ertu; Taylor, Andrew

    2010-01-01

    This presentation is given at the Virtual Heritage 2010, Bangor, UK. The team is invited to present the progress of the current research & development in the 3D Digital Heritage visualisation project based on UNESCO world Heritage site at Saltaire in West Yorkshire UK.

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

    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\\...

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

    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.

  20. Photopatterning of Hydrogel Scaffolds Coupled to Filter Materials Using Stereolithography for Perfused 3D Culture of Hepatocytes

    Shepard Neiman, Jaclyn A.; 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.

    2015-01-01

    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 fabricate...

  1. Influence of electrical stimulation on 3D-cultures of Adipose Tissue Derived Progenitor Cells (ATDPCs) behavior

    Castells-Sala, C.; Sánchez Terrones, Benjamín; Recha-Sancho, L.; V. Puig; Bragós Bardia, Ramon; C. E. Semino

    2012-01-01

    Tissue engineering has a fundamental role in regenerative medicine. Still today, the major motivation for cardiac regeneration is to design a platform that enables the complete tissue structure and physiological function regeneration of injured myocardium areas. Although tissue engineering approaches have been generally developed for two-dimensional (2D) culture systems, three-dimensional (3D) systems are being spotlighted as the means to mimic better in vivo cellular conditions. This manuscr...

  2. State-of-the-Art of 3D Cultures (Organs-on-a-Chip) in Safety Testing and Pathophysiology

    Alepee, Natalie; Bahinski, Anthony; Daneshian, Mardas; De Weyer, Bart; Fritsche, Ellen; Goldberg, Alan; Hansmann, Jan; Hartung, Thomas; Haycock, John; Hogberg, Helena T.; Hoelting, Lisa; Jens M Kelm; Kadereit, Suzanne; McVey, Emily; Landsiedel, Robert

    2015-01-01

    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 ...

  3. Increased GFAP immunoreactivity by astrocytes in response to contact with dorsal root ganglia cells in a 3D culture model

    East, Emma; Golding, Jon; Phillips, James

    2007-01-01

    Failure of repair mechanisms in the injured CNS is widely attributed to the inhibitory environment of the lesion site, most notably the formation of the glial scar which forms a physical and physiological barrier to axon regeneration. We developed an in vitro 3D cell culture model to investigate the response of astrocytes to cells found at the inhibitory interfaces formed following damage to the spinal cord. CellTrackerTM labelled dissociated DRGs were seeded onto astrocy...

  4. Design, construction and testing of a COC 3D flow-over flow-through bioreactor for hepatic cell culture

    Windels, Jindrich; Verplancke, Rik; Jahanshahi, Amir; Heimann, Marcus; Leite, Sofia B.; Roosens, Tiffany; van Grunsven, Leo A; Barbe, Laurent; Prill, Sebastian; Jaeger, Magnus; Duschl, Claus; Vanfleteren, Jan

    2015-01-01

    In this poster, we present the joint development efforts for a 3D microfluidic bioreactor for hepatic cell cultures. Cyclic Olefin Copolymer (COC) was selected for constructing the bioreactor, since the material has good chemical resistance, low adsorption and good optical properties, including low auto-fluorescence. A downside of COC is that it is much more difficult to structure than more traditional microfluidic materials, such as PDMS, PMMA, … Two parallel approaches were developed for...

  5. Advances in 3D Spatial Information Systems. Applications in cultural heritage and virtual archeology

    María Dolores Robles Ortega

    2015-10-01

    Full Text Available Large point clouds from radars and these-dimensional scanners are commonly used in Archaeology. However, in most cases these models cannot be properly integrated and used in software such as heritage management due to its large size. Therefore, some tools to make this management easier and optimize the processing are needed. In this work, we propone the integration between OpenVDB and GRASS in a C++ module to combine the widen functionality of GRASS GIS with the 3D models management efficiency of OpenVDB. Specifically, this application is used to combine the topographic information of a city with the 3D models of the most significant buildings. This application can be useful for both current cities as well as for virtual reconstruction of existing villages in the olden days and currently disappeared.

  6. 3D Visualization of Cultural Heritage Artefacts with Virtual Reality devices

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

    2015-01-01

    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....

  7. Virtual 3D museum of Neolithic culture. Artefacts, technology and imagining

    Květina, Petr; Brzobohatá, Hana; Burgert, Pavel; Končelová, Markéta; Pavlů, Ivan; Unger, Jiří; Řídký, Jaroslav; Šumberová, Radka; Vavrečka, Petr

    Istanbul: Archaeology & Art Publications, 2014 - (Yilmaz, Ö.). s. 205-206 ISBN 978-605-396-287-8. [Annual Meeting of the European Association of Archaeologists /20./. 10.09.2014-14.09.2014, Istanbul] R&D Projects: GA MK(CZ) DF12P01OVV032 Keywords : Neolithic * virtual museum * 3D * augmented reality Subject RIV: AC - Archeology, Anthropology, Ethnology https://www.eaa2014istanbul.org/assets/indirilecekler/2014%20EAA%20abstracts.pdf

  8. Assessments for 3d Reconstructions of Cultural Heritage Using Digital Technologies

    Manferdini, A. M.; Galassi, M.

    2013-02-01

    The aim of this contribution is to show the results of evaluations on 3D digitizations performed using different methodologies and technologies. In particular, for surveys conducted at the architectural and urban scale, the recent reduction of costs related to Time of Flight and phase shift laser scanners is actually enhancing the replacement of traditional topographic instruments (i.e. total stations) with range-based technologies for the acquisition of 3D data related to built heritage. If compared to surveys performed using traditional topographic technologies, range-based ones offer a wide range of advantages, but they also require different skills, procedures and times. The present contribution shows the results of a practical application of both approaches on the same case study. Another application was suggested by the recent developments in the photogrammetric field that enhance the improvement of software able to automatically orient uncalibrated cameras and derive dense and accurate 3D point clouds, with evident benefits in reduction of costs required for survey equipment. Therefore, the presented case study constituted the occasion to compare a rangebased survey with a fast 3D acquisition and modelling using a Structure from Motion solution. These survey procedures were adopted at an architectural scale, on a single building, that was surveyed both on the outside and on the inside. Assessments on the quality of the rebuilt information is reported, as far as metric accuracy and reliability is concerned, as well as on time consuming and on skills required during each step of the adopted pipelines. For all approaches, these analysis highlighted advantages and disadvantages that allow to conduct evaluations on the possible convenience of adopting range-based technologies instead of a traditional topographic approach or a photogrammetric one instead of a range based one in case of surveys conducted at an architectural/urban scale.

  9. Microtubule regulation of corneal fibroblast morphology and mechanical activity in 3-D culture

    Kim, Areum; Petroll, W. Matthew

    2007-01-01

    The purpose of this study was to investigate the role of microtubules in regulating corneal fibroblast structure and mechanical behavior using static (3-D) and dynamic (4-D) imaging of both cells and their surrounding matrix. Human corneal fibroblasts transfected to express GFP-zyxin (to label focal adhesions) or GFP-tubulin (to label microtubules) were plated at low density inside 100 μm thick type I collagen matrices. After 24 hours, the effects of nocodazole (to depolymerize microtubules),...

  10. The Effects of GDF-5 and Uniaxial Strain on Mesenchymal Stem Cells in 3-D Culture

    Farng, Eugene; Urdaneta, Alfonso R.; Barba, David; Esmende, Sean; McAllister, David R.

    2008-01-01

    Recent endeavors in tissue engineering have attempted to identify the optimal parameters to create an artificial ligament. Both mechanical and biochemical stimulation have been used by others to independently modulate growth and differentiation, although few studies have explored their interactions. We applied previously described fabrication techniques to create a highly porous (90%–95% porosity, 212–300 μm), 3-D, bioabsorbable polymer scaffold (polycaprolactone). Scaffolds were coated with ...

  11. Mechanisms of DNA damage response to targeted irradiation in organotypic 3D skin cultures.

    Anna Acheva

    Full Text Available DNA damage (caused by direct cellular exposure and bystander signaling and the complex pathways involved in its repair are critical events underpinning cellular and tissue response following radiation exposures. There are limited data addressing the dynamics of DNA damage induction and repair in the skin particularly in areas not directly exposed. Here we investigate the mechanisms regulating DNA damage, repair, intracellular signalling and their impact on premature differentiation and development of inflammatory-like response in the irradiated and surrounding areas of a 3D organotypic skin model. Following localized low-LET irradiation (225 kVp X-rays, low levels of 53BP1 foci were observed in the 3D model (3.8±0.28 foci/Gy/cell with foci persisting and increasing in size up to 48 h post irradiation. In contrast, in cell monolayers 14.2±0.6 foci/Gy/cell and biphasic repair kinetics with repair completed before 24 h was observed. These differences are linked to differences in cellular status with variable level of p21 driving apoptotic signalling in 2D and accelerated differentiation in both the directly irradiated and bystander areas of the 3D model. The signalling pathways utilized by irradiated keratinocytes to induce DNA damage in non-exposed areas of the skin involved the NF-κB transcription factor and its downstream target COX-2.

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

    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

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

    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.

  14. Response of the Gulf of Alaska 3D winter circulation to oceanic climate shifts: Ecosystem implications

    Auad, Guillermo

    2008-01-01

    Recent measurements of zooplankton biomass and biological productivity in the Gulf of Alaska have raised a number of questions regarding possible linkages between climate and availability of renewable resources. In this article we compare 3 abrupt oceanic regime shifts in the Gulf of Alaska, the 1976-1977 warming shift, the 1998 cooling episode, and the 1998 to 1999 El Niño to La Niña transition, against concomitant changes in biological conditions reported in the literature. After the 1976-1977 warming shift, changes in the Gulf's 3D circulation, i.e., interior upwelling, onshore transport and coastal downwelling, had the same sign as their climatological means, thus providing a candidate explanation to the observed increased productivity of the upper ocean. Warming and cooling transitions have associated with them very different patterns of both horizontal and vertical circulation, where the latter is confirmed to be linked to the wind stress curl variability. Important shifts in the local biology have been reported in the literature (warming of 1976-1997 and El Niño to La Niña transition of 1998-1999) when climatological shifts in our simulated vertical velocities were large; in turn, when they were small, the ecosystem did not show significant changes and this was in part due to the resilience set by the 1976-1977 shift through the strengthening of the GOA's mean 3D circulation.

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

    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

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

    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.

  17. Establishment of 3D Co-Culture Models from Different Stages of Human Tongue Tumorigenesis: Utility in Understanding Neoplastic Progression

    Sawant, Sharada; Dongre, Harsh; Singh, Archana Kumari; Joshi, Shriya; Costea, Daniela Elena; Mahadik, Snehal; Ahire, Chetan; Makani, Vidhi; Dange, Prerana; Sharma, Shilpi; Chaukar, Devendra; Vaidya, Milind

    2016-01-01

    To study multistep tumorigenesis process, there is a need of in-vitro 3D model simulating in-vivo tissue. Present study aimed to reconstitute in-vitro tissue models comprising various stages of neoplastic progression of tongue tumorigenesis and to evaluate the utility of these models to investigate the role of stromal fibroblasts in maintenance of desmosomal anchoring junctions using transmission electron microscopy. We reconstituted in-vitro models representing normal, dysplastic, and malignant tissues by seeding primary keratinocytes on either fibroblast embedded in collagen matrix or plain collagen matrix in growth factor-free medium. The findings of histomorphometry, immunohistochemistry, and electron microscopy analyses of the three types of 3D cultures showed that the stratified growth, cell proliferation, and differentiation were comparable between co-cultures and their respective native tissues; however, they largely differed in cultures grown without fibroblasts. The immunostaining intensity of proteins, viz., desmoplakin, desmoglein, and plakoglobin, was reduced as the disease stage increased in all co-cultures as observed in respective native tissues. Desmosome-like structures were identified using immunogold labeling in these cultures. Moreover, electron microscopic observations revealed that the desmosome number and their length were significantly reduced and intercellular spaces were increased in cultures grown without fibroblasts when compared with their co-culture counterparts. Our results showed that the major steps of tongue tumorigenesis can be reproduced in-vitro. Stromal fibroblasts play a role in regulation of epithelial thickness, cell proliferation, differentiation, and maintenance of desmosomalanchoring junctions in in-vitro grown tissues. The reconstituted co-culture models could help to answer various biological questions especially related to tongue tumorigenesis. PMID:27501241

  18. Comparison of the Expression of Hepatic Genes by Human Wharton’s Jelly Mesenchymal Stem Cells Cultured in 2D and 3D Collagen Culture Systems

    Khodabandeh, Zahra; Vojdani, Zahra; Talaei-Khozani, Tahereh; Jaberipour, Mansoureh; Hosseini, Ahmad; Bahmanpour, Soghra

    2016-01-01

    Background: Human Wharton’s jelly mesenchymal stem cells (HWJMSCs) express liver-specific markers such as albumin, alpha-fetoprotein, cytokeratin-19, cytokeratin-18, and glucose-6-phosphatase. Therefore, they can be considered as a good source for cell replacement therapy for liver diseases. This study aimed to evaluate the effects of various culture systems on the hepatocyte-specific gene expression pattern of naïve HWJMSCs. Methods: HWJMSCs were characterized as MSCs by detecting the surface CD markers and capability to differentiate toward osteoblast and adipocyte. HWJMSCs were cultured in 2D collagen films and 3D collagen scaffolds for 21 days and were compared to control cultures. Real time RT-PCR was used to evaluate the expression of liver-specific genes. Results: The HWJMSCs which were grown on non-coated culture plates expressed cytokeratin-18 and -19, alpha-fetoprotein, albumin, glucose-6-phosphatase, and claudin. The expression of the hepatic nuclear factor 4 (HNF4) was very low. The cells showed a significant increase in caludin expression when they cultured in 3D collagen scaffolds compared to the conventional monolayer culture and 2D collagen scaffold. Conclusion: Various culture systems did not influence on hepatocyte specific marker expression by HWJMSCs, except for claudin. The expression of claudin showed that 3D collagen scaffold provided the extracellular matrix for induction of the cells to interconnect with each other. PMID:26722142

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

    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.

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

    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

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

    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

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

    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.

  3. Effect of Aflatoxin B1 on Growth of Bovine Mammary Epithelial Cells in 3D and Monolayer Culture System

    Babak Qasemi-Panahi

    2013-02-01

    Full Text Available Purpose: Many studies have been showed transfer of aflatoxins, toxins produced by Aspergillus flvaus and Aspergillus parasiticus fungi, into milk. These toxins are transferred into the milk through digestive system by eating contaminated food. Due to the toxicity of these materials, it seems that it has side effects on the growth of mammary cells. Therefore, the present work aimed to investigate possible toxic effects of aflatoxin B1 (AFB1 on bovine mammary epithelial cells in monolayer and three-dimensional cultures. Methods: Specimens of the mammary tissue of bovine were sized out in size 2×2 cm in slaughterhouse. After disinfection and washing in sterile PBS, primary cell culture was performed by enzymatic digestion of tissue with collagenase. When proper numbers of cells were achieved in monolayer culture, cells were seeded in a 24-well culture plate for three-dimensional (3D culture in Matrigel matrix. After 21 days of 3D culture and reaching the required number of cells, the concentrations of 15, 25 and 35 μL of AFB1 were added to the culture in quadruplicate and incubated for 8 hours. Cellular cytotoxicity was examined using standard colorimetric assay and finally, any change in the morphology of the cells was studied by microscopic technique. Results: Microscopic investigations showed necrosis of the AFB1-exposed cells compared to the control cells. Also, bovine mammary epithelial cells were significantly affected by AFB1 in dose and time dependent manner in cell viability assays. Conclusion: According to the results, it seems that AFB1 can induce cytotoxicity and necrosis in bovine mammary epithelial cells.

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

    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).

  5. Probabilistic Seismic Hazard Maps of Seattle, Washington, Including 3D Sedimentary Basin Effects and Rupture Directivity: Implications of 3D Random Velocity Variations (Invited)

    Frankel, A. D.; Stephenson, W. J.; Carver, D.; Odum, J.; Williams, R. A.; Rhea, S.

    2010-12-01

    We have produced probabilistic seismic hazard maps of Seattle for 1 Hz spectral acceleration, using over five hundred 3D finite-difference simulations of earthquakes on the Seattle fault, Southern Whidbey Island fault, and Cascadia subduction zone, as well as for random deep and shallow earthquakes at various locations. The 3D velocity model was validated by modeling the observed waveforms for the 2001 M6.8 Nisqually earthquake and several smaller events in the region. At these longer periods (≥ 1 sec) that are especially important to the response of buildings of ten stories or higher, seismic waves are strongly influenced by sedimentary basins and rupture directivity. We are investigating how random spatial variations in the 3D velocity model affect the simulated ground motions for M6.7 earthquakes on the Seattle fault. A fractal random variation of shear-wave velocity with a Von Karman correlation function produces spatial variations of peak ground velocity with multiple scale lengths. We find that a 3D velocity model with a 10% standard deviation in shear-wave velocity in the top 1.5 km and 5% standard deviation from 1.5-10 km depth produces variations in peak ground velocities of as much as a factor of two, relative to the case with no random variations. The model with random variations generally reduces the peak ground velocity of the forward rupture directivity pulse for sites near the fault where basin-edge focusing of S-waves occurs. It also tends to reduce the peak velocity of localized areas where basin surface waves are focused. However, the medium with random variations also causes small-scale amplification of ground motions over distances of a few kilometers. We are also evaluating alternative methods of characterizing the aleatory uncertainty in the probabilistic hazard calculations.

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

    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

  7. Optimization of a 3D Dynamic Culturing System for In Vitro Modeling of Frontotemporal Neurodegeneration-Relevant Pathologic Features

    Tunesi, Marta; Fusco, Federica; Fiordaliso, Fabio; Corbelli, Alessandro; Biella, Gloria; Raimondi, Manuela T.

    2016-01-01

    Frontotemporal lobar degeneration (FTLD) is a severe neurodegenerative disorder that is diagnosed with increasing frequency in clinical setting. Currently, no therapy is available and in addition the molecular basis of the disease are far from being elucidated. Consequently, it is of pivotal importance to develop reliable and cost-effective in vitro models for basic research purposes and drug screening. To this respect, recent results in the field of Alzheimer’s disease have suggested that a tridimensional (3D) environment is an added value to better model key pathologic features of the disease. Here, we have tried to add complexity to the 3D cell culturing concept by using a microfluidic bioreactor, where cells are cultured under a continuous flow of medium, thus mimicking the interstitial fluid movement that actually perfuses the body tissues, including the brain. We have implemented this model using a neuronal-like cell line (SH-SY5Y), a widely exploited cell model for neurodegenerative disorders that shows some basic features relevant for FTLD modeling, such as the release of the FTLD-related protein progranulin (PRGN) in specific vesicles (exosomes). We have efficiently seeded the cells on 3D scaffolds, optimized a disease-relevant oxidative stress experiment (by targeting mitochondrial function that is one of the possible FTLD-involved pathological mechanisms) and evaluated cell metabolic activity in dynamic culture in comparison to static conditions, finding that SH-SY5Y cells cultured in 3D scaffold are susceptible to the oxidative damage triggered by a mitochondrial-targeting toxin (6-OHDA) and that the same cells cultured in dynamic conditions kept their basic capacity to secrete PRGN in exosomes once recovered from the bioreactor and plated in standard 2D conditions. We think that a further improvement of our microfluidic system may help in providing a full device where assessing basic FTLD-related features (including PRGN dynamic secretion) that may

  8. 3D SURVEY AND AUGMENTED REALITY FOR CULTURAL HERITAGE. THE CASE STUDY OF AURELIAN WALL AT CASTRA PRAETORIA IN ROME

    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

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

    Sunita Nayak

    Full Text Available 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.

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

    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...

  11. 3D modeling of GJ1214b's atmosphere: formation of inhomogeneous high clouds and observational implications

    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...

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

    Wrzesinski, Krzysztof; Fey, S. J.

    2011-01-01

    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...

  13. 3D airflow dynamics over transverse ridges Mpekweni, South Africa: implications for dune field migration behaviour

    Jackson, Derek; Cooper, Andrew; Green, Andrew; Beyers, Meiring; Wiles, Errol; Benallack, Keegan

    2016-04-01

    Un-vegetated dune fields provide excellent opportunities to examine airflow dynamics over various types and scales of dune landforms. The three dimensional surface over which lower boundary layers travel, help adjust surface airflow and consequently the aeolian response of the dunes themselves. The use of computational fluid dynamic (CFD) modelling in recent studies now enables investigation of the 3D behaviour of airflow over complex terrain, providing new insights into heterogeneous surface flow and aeolian response of dune surfaces on a large (dunefield) scale. Using a largely un-vegetated coastal dune field site at Mpekweni, Eastern Cape, South Africa, a detailed (0.1m gridded) terrestrial laser scanning survey was conducted to create a high resolution topographical surface. Using local wind flow measurements and local met station records as input, CFD modelling was performed for a number of scenarios involving variable direction and magnitude to examine surface flow patterns across multiple dune forms. Near surface acceleration, expansion and separation of airflow inducing convergence and divergence (steering) of flow velocity streamlines are investigated. Flow acceleration over dune crests/brink lines is a key parameter in driving dune migration and slip face dynamics. Dune aspect ratio (height to length) is also important in determining the degree of crestal flow acceleration, with an increase in flow associated with increasing aspect ratios. Variations in dune height appear to be the most important parameter in driving general flow acceleration. The results from the study provide new insights into dune migration behaviour at this site as well as surface flow behaviour across multiple dune configurations and length scales within un-vegetated dune fields.

  14. Comparison and functional implications of the 3D architectures of viral tRNA-like structures.

    Hammond, John A; Rambo, Robert P; Filbin, Megan E; Kieft, Jeffrey S

    2009-02-01

    RNA viruses co-opt the host cell's biological machinery, and their infection strategies often depend on specific structures in the viral genomic RNA. Examples are tRNA-like structures (TLSs), found at the 3' end of certain plant viral RNAs, which can use the cell's aminoacyl tRNA-synthetases (AARSs) to drive addition of an amino acid to the 3' end of the viral RNA. TLSs are multifunctional RNAs involved in processes such as viral replication, translation, and viral RNA stability; these functions depend on their fold. Experimental result-based structural models of TLSs have been published. In this study, we further examine these structures using a combination of biophysical and biochemical approaches to explore the three-dimensional (3D) architectures of TLSs from the turnip yellow mosaic virus (TYMV), tobacco mosaic virus (TMV), and brome mosaic virus (BMV). We find that despite similar function, these RNAs are biophysically diverse: the TYMV TLS adopts a characteristic tRNA-like L shape, the BMV TLS has a large compact globular domain with several helical extensions, and the TMV TLS aggregates in solution. Both the TYMV and BMV TLS RNAs adopt structures with tight backbone packing and also with dynamic structural elements, suggesting complexities and subtleties that cannot be explained by simple tRNA mimicry. These results confirm some aspects of existing models and also indicate how these models can be improved. The biophysical characteristics of these TLSs show how these multifunctional RNAs might regulate various viral processes, including negative strand synthesis, and also allow comparison with other structured RNAs. PMID:19144910

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

    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.

  16. Using LIDO to handle 3D cultural heritage documentation data provenance

    Pitzalis, Denis; Niccolucci, Franco; Cord, Matthieu

    2011-01-01

    International audience It is important for Digital Libraries (DL) to be flexible in exposing their content. Typically a DL provides a search/browse interface which allows resources to be found and a service to make the data available for harvesting from/to other DLs. This kind of communication is possible because the structures of different DLs are expressed following formal specifications. In particular in Cultural Heritage, where we need to describe an extremely heterogeneous environment...

  17. Assessment of different 3D culture systems to study tumor phenotype and chemosensitivity in pancreatic ductal adenocarcinoma.

    Zeeberg, Katrine; Cardone, Rosa Angela; Greco, Maria Raffaella; Saccomano, Mara; Nøhr-Nielsen, Asbjørn; Alves, Frauke; Pedersen, Stine Falsig; Reshkin, Stephan Joel

    2016-07-01

    Pancreatic ductal adenocarcinoma (PDAC) is a highly malignant disease with a very poor prognosis, due to the influence of the tumor stroma, which promotes tumor growth, early invasion and chemoradiation resistance. Efforts to develop models for identifying novel anticancer therapeutic compounds have been hampered by the limited ability of in vitro models to mimic these in vivo tumor-stroma interactions. This has led to the development of various three-dimensional (3D) culture platforms recapitulating the in vivo tumor-stroma crosstalk and designed to better understand basic cancer processes and screen drug action. However, a consensus for different experimental 3D platforms is still missing in PDAC. We compared four PDAC cell lines of different malignancy grown in 2D monolayers to three of the more commonly used 3D techniques (ultralow adhesion concave microwells, Matrigel inclusion and organotypic systems) and to tumors derived from their orthotopic implantation in mice. In these 3D platforms, we observed that cells grow with very different tumor morphologies and the organotypic setting most closely resembles the tumor cytoarchitecture obtained by orthotopically implanting the four cell lines in mice. We then analyzed the molecular and cellular responses of one of these cell lines to epidermal growth factor receptor (EGFR) stimulation with EGF and inhibition with erlotinib and found that only in the 3D platforms, and especially the organotypic, cells: i) responded to EGF by changing the expression of signalling components underlying cell-stroma crosstalk and tissue architecture, growth, invasion and drug resistance (E-cadherin, EGFR, ezrin, β1 integrin, NHERF1 and HIF-1α) similar to those reported in vivo; ii) had stimulated growth and increased erlotinib sensitivity in response to EGF, more faithfully mimicking their known in vivo behaviour. Altogether, these results, indicate the organotypic as the most relevant physiological 3D system to study the

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

    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

  19. New insights on 3-D plates interaction near Taiwan from tomography and tectonic implications

    Lallemand, Serge; Font, Yvonne; Bijwaard, Harmen; Kao, Honn

    2001-07-01

    Recent tomographic results are used to trace the South China Sea and Philippine Sea subducting slabs, south and northeast of Taiwan along the Manila and Ryukyu trenches, respectively. In particular, the 3-D plates interaction beneath Taiwan is discussed based on a close-up view of the tomographic sections together with earthquake hypocenters distribution. Our study indicates that: (1) the east-dipping South China Sea slab can be followed to the north, up to the latitude of Hualien, (2) the Eurasian plate subducts beneath most part of the Taiwan island down to the 670 km-depth discontinuity, (3) the north-dipping Philippine Sea slab can be followed slightly west of the longitude of Hualien. Both plates thus interact beneath northern Taiwan where the arc-continent collision is paroxysmal. (4) Slab detachment is envisaged at the northern edge of the subducted Eurasian plate beneath the Coastal Range of Taiwan, which may facilitate the northwestward motion of the Philippine Sea plate with respect to Eurasia. Slabs geometries obtained from tomographic sections allow us to reconstruct the Late Neogene plate kinematics and dynamics in this region. Our main conclusions are: (1) The size of the original South China Sea was about twice its present size. (2) The subducted part of the West Philippine Basin i.e. the largest oceanic basin of the Philippine Sea Plate, extends only 400 km north of the Ryukyu Trench. (3) Slab detachment might have occurred 3-5 my ago beneath the central and northern Ryukyu Arc along a weak zone that is aligned with the Gagua Ridge: an ancient plate boundary. (4) The Ryukyu Trench has propagated westward from 126°E of longitude (southeast of Miyako Island) to the locus of the present arc-continent collision, along a major lithospheric tear that cut through the continent-ocean boundary first, and then through the continental lithosphere. As a consequence, the southern Ryukyu margin developed progressively from east to west as a subduction zone

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

    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.

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

    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. PMID:25962333

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

    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 μm 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

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

    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

  4. [Novel methods for studies of testicular development and spermatogenesis: From 2D to 3D culture].

    Zhang, Lian-dong; Li, He-cheng; Zhang, Tong-dian; Wang, Zi-ming

    2016-03-01

    The two-dimensional model of cell culture is an important method in the study of testicular development and spermatogenesis but can not effectively mimic and regulate the testicular microenvironment and the whole process of spermatogenesis due to the lack of relevant cell factors and the disruption of a three-dimensional spatial structure. In the past 20 years, the development and optimization of the in vitro model such as testis organotypic culture and in vivo model such as testis transplantation achieved a transformation from two- to three-dimension. The maintenance and optimization of the testicular niche structure could mimic the testicular microenvironment and cell types including Leydig, Sertoli and germ cells, which showed similar biological behaviors to those in vivo. Besides, the cell suspension or tissue fragment floats in the gas-liquid interface so that the development of somatic and germ cells is well maintained in vitro whilst the feedback linkage between grafted testis tissue and hypothalamus-pituitary of the host rebuilt in the in vitro model provides an endocrinological basis for spermatogenesis, which serves as an effective methodology to better understand the organogenesis and development of the testis as well as testicular function regulation, advancing the concept of treatment of male infertility. Al- though each of the methods may have its limitations, the progress in the processing, freezing, thawing, and transplantation of cells and tissues will surely promote their clinical application and present their value in translational medicine. PMID:27172668

  5. 3DMADMAC|SPECTRAL: Hardware and Software Solution for Integrated Digitization of 3D Shape, Multispectral Color and BRDF for Cultural Heritage Documentation

    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.

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

    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). PMID:21884222

  7. Establishment of 3D organotypic cultures using human neonatal epidermal cells.

    Gangatirkar, Pradnya; Paquet-Fifield, Sophie; Li, Amy; Rossi, Ralph; Kaur, Pritinder

    2007-01-01

    This protocol describes an ex vivo three-dimensional coculture system optimized to study the skin regenerative ability of primary human keratinocytes grown at the air-liquid interface on collagen matrices embedded with human dermal fibroblasts. An option for enrichment of keratinocyte stem cells and their progeny using fluorescence-activated cell sorting is also provided. Initially, dermal equivalents, comprising human passaged fibroblasts seeded in a collagen matrix, are grown on porous filters (3 mum) placed in transwells. After 1 week, primary human keratinocytes are seeded on this base. One week later, an air-lift transition is performed, leading to the differentiation of the keratinocytes, which are macroscopically visible as artificial skin after a couple of days. The cultures can be harvested 1 week after the air-lift and processed for immunohistochemistry or gene expression analysis. The overall procedure can be completed in 3 weeks, including the preparation of the dermal equivalent and the seeding of the primary keratinocytes. PMID:17401352

  8. 3D Culture of Chondrocytes in Gelatin Hydrogels with Different Stiffness

    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.

  9. Effectiveness of octenidine and chlorhexidine in the artificially contaminated 3-D-culture of human keratinocytes

    Kramer, Axel

    2009-12-01

    Full Text Available Epidermis-Equivalents (EpiDerm derived from human keratinocytes were obtained after 14 d air-lift-culture. These In-vitro-models were used for the demonstration of a possible postantiseptic effect against the test microorganisms Pseudomonas aeruginosa and Staphylococcus aureus after 5 min pre-incubation of the surface of the epidermis with equimolar concentrations of the antiseptic agents chlorhexidine digluconate (CHX and octenidine dihydrochloride (OCT of 1.6 mmol/l. Both antiseptics adhere to the surface of EpiDerm. Each active agent cannot be removed by washing with phosphate-buffered saline (PBS and remains microbiocidal active. EpiDerm-OCT was more effective after 30 min at room temperature against 106 cfu/ml Staphylococcus aureus resulting 3.2–4 log10 reduction, but not for EpiDerm-CHX producing only 2–2.4 log10 reduction. A maximum of 0.6–1.2 log10 reduction was found against Pseudomonas aeruginosa after 30 min exposure in EpiDerm-OCT, but there was no or a neglectable microbiocidal activity of 0–0.2 log10 reduction using EpiDerm-CHX. The postantiseptic effect of OCT is superior to EpiDerm-bound CHX comparing equimolar concentrations of active agent used for pre-incubation of EpiDerm. Treatment of EpiDerm with OCT alone or in combination with test organisms resulted in no cytotoxic effect in viable keratinocytes. In contrast to that the test organism Staphylococcus aureus, the active agent CHX alone or in combination with both test organisms demonstrated a cytotoxic activity reducing the viability of basal keratinocytes more than 10%. Therefore, OCT is more tolerated than CHX by basal keratinocytes of the presented In-vitro-model EpiDerm.

  10. The role of 3D microenvironmental organization in MCF-7 epithelial–mesenchymal transition after 7 culture days

    Foroni, Laura [Pathology Unit, Department of Haematology, Oncology and Clinical Pathology, S. Orsola-Malpighi Hospital, Bologna University (Italy); Vasuri, Francesco, E-mail: vasurifrancesco@libero.it [Pathology Unit, Department of Haematology, Oncology and Clinical Pathology, S. Orsola-Malpighi Hospital, Bologna University (Italy); Chair of Vascular Surgery, Department of Specialistic Surgery and Anaesthesiological Sciences, S. Orsola-Malpighi Hospital, Bologna University (Italy); Valente, Sabrina [Pathology Unit, Department of Haematology, Oncology and Clinical Pathology, S. Orsola-Malpighi Hospital, Bologna University (Italy); Gualandi, Chiara [Department of Chemistry “G. Ciamician” and National Consortium of Materials Science and Technology (INSTM, RU Bologna), Bologna University (Italy); Focarete, Maria Letizia [Department of Chemistry “G. Ciamician” and National Consortium of Materials Science and Technology (INSTM, RU Bologna), Bologna University (Italy); Health Science and Technologies–Interdepartmental Center for Industrial Research (HST-ICIR), Bologna University (Italy); Caprara, Giacomo [Pathology Unit, Department of Haematology, Oncology and Clinical Pathology, S. Orsola-Malpighi Hospital, Bologna University (Italy); Scandola, Mariastella [Department of Chemistry “G. Ciamician” and National Consortium of Materials Science and Technology (INSTM, RU Bologna), Bologna University (Italy); D' Errico-Grigioni, Antonia; Pasquinelli, Gianandrea [Pathology Unit, Department of Haematology, Oncology and Clinical Pathology, S. Orsola-Malpighi Hospital, Bologna University (Italy)

    2013-06-10

    We present a multi-technique study on in vitro epithelial–mesenchymal transition (EMT) in human MCF-7 cells cultured on electrospun scaffolds of poly(L-lactic acid) (PLA), with random and aligned fiber orientations. Our aim is to investigate the morphological and genetic characteristics induced by extracellular matrix in tumor cells cultured in different 3D environments, and at different time points. Cell vitality was assessed with AlamarBlue at days 1, 3, 5 and 7. Scanning electron microscopy was performed at culture days 3 and 7. Immunohistochemistry (for E-cadherin, β-catenin, cytokeratins, nucleophosmin, tubulin, Ki-67 and vimentin), immunofluorescence (for F-actin) western blot (for E-cadherin, β-catenin and vimentin) and transmission electron microscopy were carried out at day 7. An EMT gene array followed by PCR analysis confirmed the regulation of selected genes. At day 7, scanning electron microscopy on aligned-PLA revealed spindle-shaped cells gathered in buds and ribbon-like structures, with a higher nucleolar/nuclear ratio and a loss in E-cadherin and β-catenin at immunohistochemistry and western blot. An up-regulation of SMAD2, TGF-β2, TFPI2 and SOX10 was found in aligned-PLA compared to random-PLA cultured cells. The topography of the extracellular matrix has a role in tumor EMT, and a more aggressive phenotype characterizes MCF-7 cells cultured on aligned-PLA scaffold. -- Highlights: • After 7 culture days an aligned-PLA scaffold induces a spindle shape to MCF-7 cells. • Despite these changes, the aligned MCF-7 cells keep an epithelial phenotype. • The extracellular environment alone influences the E-cadherin/β-catenin axis. • The extracellular environment can promote the epithelial–mesenchymal transition.

  11. The role of 3D microenvironmental organization in MCF-7 epithelial–mesenchymal transition after 7 culture days

    We present a multi-technique study on in vitro epithelial–mesenchymal transition (EMT) in human MCF-7 cells cultured on electrospun scaffolds of poly(L-lactic acid) (PLA), with random and aligned fiber orientations. Our aim is to investigate the morphological and genetic characteristics induced by extracellular matrix in tumor cells cultured in different 3D environments, and at different time points. Cell vitality was assessed with AlamarBlue at days 1, 3, 5 and 7. Scanning electron microscopy was performed at culture days 3 and 7. Immunohistochemistry (for E-cadherin, β-catenin, cytokeratins, nucleophosmin, tubulin, Ki-67 and vimentin), immunofluorescence (for F-actin) western blot (for E-cadherin, β-catenin and vimentin) and transmission electron microscopy were carried out at day 7. An EMT gene array followed by PCR analysis confirmed the regulation of selected genes. At day 7, scanning electron microscopy on aligned-PLA revealed spindle-shaped cells gathered in buds and ribbon-like structures, with a higher nucleolar/nuclear ratio and a loss in E-cadherin and β-catenin at immunohistochemistry and western blot. An up-regulation of SMAD2, TGF-β2, TFPI2 and SOX10 was found in aligned-PLA compared to random-PLA cultured cells. The topography of the extracellular matrix has a role in tumor EMT, and a more aggressive phenotype characterizes MCF-7 cells cultured on aligned-PLA scaffold. -- Highlights: • After 7 culture days an aligned-PLA scaffold induces a spindle shape to MCF-7 cells. • Despite these changes, the aligned MCF-7 cells keep an epithelial phenotype. • The extracellular environment alone influences the E-cadherin/β-catenin axis. • The extracellular environment can promote the epithelial–mesenchymal transition

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

    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.

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

    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.

  14. Metabolic response of lung cancer cells to radiation in a paper-based 3D cell culture system.

    Simon, Karen A; Mosadegh, Bobak; Minn, Kyaw Thu; Lockett, Matthew R; Mohammady, Marym R; Boucher, Diane M; Hall, Amy B; Hillier, Shawn M; Udagawa, Taturo; Eustace, Brenda K; Whitesides, George M

    2016-07-01

    This work demonstrates the application of a 3D culture system-Cells-in-Gels-in-Paper (CiGiP)-in evaluating the metabolic response of lung cancer cells to ionizing radiation. The 3D tissue-like construct-prepared by stacking multiple sheets of paper containing cell-embedded hydrogels-generates a gradient of oxygen and nutrients that decreases monotonically in the stack. Separating the layers of the stack after exposure enabled analysis of the cellular response to radiation as a function of oxygen and nutrient availability; this availability is dictated by the distance between the cells and the source of oxygenated medium. As the distance between the cells and source of oxygenated media increased, cells show increased levels of hypoxia-inducible factor 1-alpha, decreased proliferation, and reduced sensitivity to ionizing radiation. Each of these cellular responses are characteristic of cancer cells observed in solid tumors. With this setup we were able to differentiate three isogenic variants of A549 cells based on their metabolic radiosensitivity; these three variants have known differences in their metastatic behavior in vivo. This system can, therefore, capture some aspects of radiosensitivity of populations of cancer cells related to mass-transport phenomenon, carry out systematic studies of radiation response in vitro that decouple effects from migration and proliferation of cells, and regulate the exposure of oxygen to subpopulations of cells in a tissue-like construct either before or after irradiation. PMID:27116031

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

    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. 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. 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. 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 induce near-complete tumor phenotype reversion. These human

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

    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 photopolymerizable hydrogel for 3-D culture of human embryonic stem cell-derived cardiomyocytes and rat neonatal cardiac cells.

    Shapira-Schweitzer, Keren; Habib, Manhal; Gepstein, Lior; Seliktar, Dror

    2009-02-01

    The purpose of this study was to assess the in vitro ability of two types of cardiomyocytes (cardiomyocytes derived from human embryonic stem cells (hESC-CM) and rat neonatal cardiomyocytes (rN-CM)) to survive and generate a functional cardiac syncytium in a three-dimensional in situ polymerizable hydrogel environment. Each cell type was cultured in a PEGylated fibrinogen (PF) hydrogel for up to two weeks while maturation and cardiac function were documented in terms of spontaneous contractile behavior and biomolecular organization. Quantitative contractile parameters including contraction amplitude and synchronization were measured by non-invasive image analysis. The rN-CM demonstrated the fastest maturation and the most significant spontaneous contraction. The hESC-CM maturation occurred between 10-14 days in culture, and exhibited less contraction amplitude and synchronization in comparison to the rN-CMs. The maturation of both cell types within the hydrogels was confirmed by cardiac-specific biomolecular markers, including alpha-sarcomeric actin, actinin, and connexin-43. Cellular responsiveness to isoproterenol, carbamylcholine and heptanol provided further evidence of the cardiac maturation in the 3-D PF hydrogel as well as identified a potential to use this system for in vitro drug screening. These findings indicate that the PF hydrogel biomaterial can be used as an in situ polymerizable biomaterial for stem cells and their cardiomyocyte derivatives. PMID:19027751

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

    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

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

    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. 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. 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. 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

  20. 3D simulation as a tool for improving the safety culture during remediation work at Andreeva Bay

    Andreeva Bay in northwest Russia hosts one of the former coastal technical bases of the Northern Fleet. Currently, this base is designated as the Andreeva Bay branch of Northwest Center for Radioactive Waste Management (SevRAO) and is a site of temporary storage (STS) for spent nuclear fuel (SNF) and other radiological waste generated during the operation and decommissioning of nuclear submarines and ships. According to an integrated expert evaluation, this site is the most dangerous nuclear facility in northwest Russia. Environmental rehabilitation of the site is currently in progress and is supported by strong international collaboration. This paper describes how the optimization principle (ALARA) has been adopted during the planning of remediation work at the Andreeva Bay STS and how Russian–Norwegian collaboration greatly contributed to ensuring the development and maintenance of a high level safety culture during this process. More specifically, this paper describes how integration of a system, specifically designed for improving the radiological safety of workers during the remediation work at Andreeva Bay, was developed in Russia. It also outlines the 3D radiological simulation and virtual reality based systems developed in Norway that have greatly facilitated effective implementation of the ALARA principle, through supporting radiological characterisation, work planning and optimization, decision making, communication between teams and with the authorities and training of field operators. (paper)

  1. 3D simulation as a tool for improving the safety culture during remediation work at Andreeva Bay.

    Chizhov, K; Sneve, M K; Szőke, I; Mazur, I; Mark, N K; Kudrin, I; Shandala, N; Simakov, A; Smith, G M; Krasnoschekov, A; Kosnikov, A; Kemsky, I; Kryuchkov, V

    2014-12-01

    Andreeva Bay in northwest Russia hosts one of the former coastal technical bases of the Northern Fleet. Currently, this base is designated as the Andreeva Bay branch of Northwest Center for Radioactive Waste Management (SevRAO) and is a site of temporary storage (STS) for spent nuclear fuel (SNF) and other radiological waste generated during the operation and decommissioning of nuclear submarines and ships. According to an integrated expert evaluation, this site is the most dangerous nuclear facility in northwest Russia. Environmental rehabilitation of the site is currently in progress and is supported by strong international collaboration. This paper describes how the optimization principle (ALARA) has been adopted during the planning of remediation work at the Andreeva Bay STS and how Russian-Norwegian collaboration greatly contributed to ensuring the development and maintenance of a high level safety culture during this process. More specifically, this paper describes how integration of a system, specifically designed for improving the radiological safety of workers during the remediation work at Andreeva Bay, was developed in Russia. It also outlines the 3D radiological simulation and virtual reality based systems developed in Norway that have greatly facilitated effective implementation of the ALARA principle, through supporting radiological characterisation, work planning and optimization, decision making, communication between teams and with the authorities and training of field operators. PMID:25254659

  2. 3D cell cultures of human head and neck squamous cell carcinoma cells are radiosensitized by the focal adhesion kinase inhibitor TAE226

    Background and purpose: Focal adhesion kinase (FAK), a main player in integrin signaling and survival, is frequently overexpressed in human cancers and therefore postulated as potential target in cancer therapy. The aim of this study was to evaluate the radiosensitizing potential of the FAK inhibitor TAE226 in three-dimensional (3D) tumor cell cultures. Materials and methods: Head and neck squamous cell carcinoma (HNSCC) cells (FaDu, UT-SCC15, UT-SCC45), lung cancer cells (A549), colorectal carcinoma cells (DLD-1, HCT-116) and pancreatic tumor cells (MiaPaCa2, Panc1) were treated with different concentrations of TAE226 (0-1 μm; 1 or 24 h) without or in combination with irradiation (0-6 Gy, X-ray, single dose). Subsequently, 3D clonogenic survival assays (laminin-rich extracellular matrix) and Western blotting (expression/phosphorylation, e.g. FAK, Akt, ERK1/2) were performed. Results: All investigated 3D cell cultures showed a dose-dependent reduction in clonogenic survival by TAE226. Intriguingly, TAE226 only significantly radiosensitized 3D HNSCC cell cultures accompanied by a pronounced dephosphorylation of FAK, Akt and ERK1/2. Conclusions: Our data demonstrate TAE226 as potent FAK inhibitor that enhances the cellular radiosensitivity particularly of HNSCC cells grown in a 3D cell culture model. Future in vitro and in vivo investigations will clarify, to which extent this approach might be clinically relevant for radiotherapy of HNSCC.

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

    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.

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

    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

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

    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

  6. Cultural Implications of Human Resource Development.

    Hiranpruk, Chaiskran

    A discussion of the cultural effects of economic and, by extension, human resource development in Southeast Asia looks at short- and long-term implications. It is suggested that in the short term, increased competition will affect distribution of wealth, which can promote materialism and corruption. The introduction of labor-saving technology may…

  7. Fluid flow increases mineralized matrix deposition in 3D perfusion culture of marrow stromal osteoblasts in a dose-dependent manner

    Bancroft, Gregory N.; Sikavitsas, Vassilios I.; van den Dolder, Juliette; Sheffield, Tiffany L.; Ambrose, Catherine G.; Jansen, John A.; Mikos, Antonios G.; McIntire, L. V. (Principal Investigator)

    2002-01-01

    Bone is a complex highly structured mechanically active 3D tissue composed of cellular and matrix elements. The true biological environment of a bone cell is thus derived from a dynamic interaction between responsively active cells experiencing mechanical forces and a continuously changing 3D matrix architecture. To investigate this phenomenon in vitro, marrow stromal osteoblasts were cultured on 3D scaffolds under flow perfusion with different rates of flow for an extended period to permit osteoblast differentiation and significant matrix production and mineralization. With all flow conditions, mineralized matrix production was dramatically increased over statically cultured constructs with the total calcium content of the cultured scaffolds increasing with increasing flow rate. Flow perfusion induced de novo tissue modeling with the formation of pore-like structures in the scaffolds and enhanced the distribution of cells and matrix throughout the scaffolds. These results represent reporting of the long-term effects of fluid flow on primary differentiating osteoblasts and indicate that fluid flow has far-reaching effects on osteoblast differentiation and phenotypic expression in vitro. Flow perfusion culture permits the generation and study of a 3D, actively modeled, mineralized matrix and can therefore be a valuable tool for both bone biology and tissue engineering.

  8. 3D co-cultures of keratinocytes and melanocytes and cytoprotective effects on keratinocytes against reactive oxygen species by insect virus-derived protein microcrystals

    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

  9. 3D co-cultures of keratinocytes and melanocytes and cytoprotective effects on keratinocytes against reactive oxygen species by insect virus-derived protein microcrystals

    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.

  10. Alginate based 3D hydrogels as an in vitro co-culture model platform for the toxicity screening of new chemical entities

    Prediction of human response to potential therapeutic drugs is through conventional methods of in vitro cell culture assays and expensive in vivo animal testing. Alternatives to animal testing require sophisticated in vitro model systems that must replicate in vivo like function for reliable testing applications. Advancements in biomaterials have enabled the development of three-dimensional (3D) cell encapsulated hydrogels as in vitro drug screening tissue model systems. In this study, we have developed an in vitro platform to enable high density 3D culture of liver cells combined with a monolayer growth of target breast cancer cell line (MCF-7) in a static environment as a representative example of screening drug compounds for hepatotoxicity and drug efficacy. Alginate hydrogels encapsulated with serial cell densities of HepG2 cells (105-108 cells/ml) are supported by a porous poly-carbonate disc platform and co-cultured with MCF-7 cells within standard cell culture plates during a 3 day study period. The clearance rates of drug transformation by HepG2 cells are measured using a coumarin based pro-drug. The platform was used to test for HepG2 cytotoxicity 50% (CT50) using commercially available drugs which further correlated well with published in vivo LD50 values. The developed test platform allowed us to evaluate drug dose concentrations to predict hepatotoxicity and its effect on the target cells. The in vitro 3D co-culture platform provides a scalable and flexible approach to test multiple-cell types in a hybrid setting within standard cell culture plates which may open up novel 3D in vitro culture techniques to screen new chemical entity compounds. - Graphical abstract: Display Omitted Highlights: → A porous support disc design to support the culture of desired cells in 3D hydrogels. → Demonstrated the co-culture of two cell types within standard cell-culture plates. → A scalable, low cost approach to toxicity screening involving multiple cell types.

  11. Printing thermoresponsive reverse molds for the creation of patterned two-component hydrogels for 3D cell culture.

    Müller, Michael; Becher, Jana; Schnabelrauch, Matthias; Zenobi-Wong, Marcy

    2013-01-01

    Bioprinting is an emerging technology that has its origins in the rapid prototyping industry. The different printing processes can be divided into contact bioprinting(1-4) (extrusion, dip pen and soft lithography), contactless bioprinting(5-7) (laser forward transfer, ink-jet deposition) and laser based techniques such as two photon photopolymerization(8). It can be used for many applications such as tissue engineering(9-13), biosensor microfabrication(14-16) and as a tool to answer basic biological questions such as influences of co-culturing of different cell types(17). Unlike common photolithographic or soft-lithographic methods, extrusion bioprinting has the advantage that it does not require a separate mask or stamp. Using CAD software, the design of the structure can quickly be changed and adjusted according to the requirements of the operator. This makes bioprinting more flexible than lithography-based approaches. Here we demonstrate the printing of a sacrificial mold to create a multi-material 3D structure using an array of pillars within a hydrogel as an example. These pillars could represent hollow structures for a vascular network or the tubes within a nerve guide conduit. The material chosen for the sacrificial mold was poloxamer 407, a thermoresponsive polymer with excellent printing properties which is liquid at 4 °C and a solid above its gelation temperature ~20 °C for 24.5% w/v solutions(18). This property allows the poloxamer-based sacrificial mold to be eluted on demand and has advantages over the slow dissolution of a solid material especially for narrow geometries. Poloxamer was printed on microscope glass slides to create the sacrificial mold. Agarose was pipetted into the mold and cooled until gelation. After elution of the poloxamer in ice cold water, the voids in the agarose mold were filled with alginate methacrylate spiked with FITC labeled fibrinogen. The filled voids were then cross-linked with UV and the construct was imaged with an

  12. Correlation between lack of norovirus replication and histo-blood group antigen expression in 3D-intestinal epithelial cultures

    Noroviruses (NoV) are a leading cause of gastroenteritis worldwide. An in vitro model for NoV replication remains elusive, making study of the virus difficult. One publication utilizing a 3-dimensional (3D) intestinal model derived from Int407 cells reported NoV replication and extensive cytopathi...

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

    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.

  14. Functional Interactions between 17β-Estradiol and Progesterone Regulate Autophagy during Acini Formation by Bovine Mammary Epithelial Cells in 3D Cultures

    Katarzyna Zielniok

    2014-01-01

    Full Text Available Mammary gland epithelium forms a network of ducts and alveolar units under control of ovarian hormones: 17-beta-estradiol (E2 and progesterone (P4. Mammary epithelial cells (MECs cultured on reconstituted basement membrane (rBM form three-dimensional (3D acini composed of polarized monolayers surrounding a lumen. Using the 3D culture of BME-UV1 bovine MECs we previously demonstrated that autophagy was induced in the centrally located cells of developing spheroids, and sex steroids increased this process. In the present study we showed that E2 and P4 enhanced the expression of ATG3, ATG5, and BECN1 genes during acini formation, and this effect was accelerated in the presence of both hormones together. The stimulatory action of E2 and P4 was also reflected by increased levels of Atg5, Atg3, and LC3-II proteins. Additionally, the activity of kinases involved in autophagy regulation, Akt, ERK, AMPK, and mTOR, was examined. E2 + P4 slightly increased the level of phosphorylated AMPK but diminished phosphorylated Akt and mTOR on day 9 of 3D culture. Thus, the synergistic actions of E2 and P4 accelerate the development of bovine mammary acini, which may be connected with stimulation of ATGs expression, as well as regulation of signaling pathways (PI3K/Akt/mTOR; AMPK/mTOR involved in autophagy induction.

  15. 3D micro-XRF for cultural heritage objects: new analysis strategies for the investigation of the Dead Sea Scrolls.

    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. PMID:21711051

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

    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.

  17. Clinical implication of parameteroptimized 3D-FISP MR angiography (MRA) in children with aortic coarctation: comparison with catheter angiography

    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.)

  18. An investigation of matching symmetry in the human pinnae with possible implications for 3D ear recognition and sound localization

    Claes, Peter; Reijniers, Jonas; Shriver, Mark D.; Snyders, Jonathan; Suetens, Paul; Nielandt, Joachim; De Tré, Guy; Vandermeulen, Dirk

    2015-01-01

    The human external ears, or pinnae, have an intriguing shape and, like most parts of the human external body, bilateral symmetry is observed between left and right. It is a well-known part of our auditory sensory system and mediates the spatial localization of incoming sounds in 3D from monaural cues due to its shape-specific filtering as well as binaural cues due to the paired bilateral locations of the left and right ears. Another less broadly appreciated aspect of the human pinna shape is ...

  19. The sequencing bias relaxed characteristics of Hi-C derived data and implications for chromatin 3D modeling.

    Peng, Cheng; Fu, Liang-Yu; Dong, Peng-Fei; Deng, Zhi-Luo; Li, Jian-Xin; Wang, Xiao-Tao; Zhang, Hong-Yu

    2013-10-01

    The 3D chromatin structure modeling by chromatin interactions derived from Hi-C experiments is significantly challenged by the intrinsic sequencing biases in these experiments. Conventional modeling methods only focus on the bias among different chromatin regions within the same experiment but neglect the bias arising from different experimental sequencing depth. We now show that the regional interaction bias is tightly coupled with the sequencing depth, and we further identify a chromatin structure parameter as the inherent characteristics of Hi-C derived data for chromatin regions. Then we present an approach for chromatin structure prediction capable of relaxing both kinds of sequencing biases by using this identified parameter. This method is validated by intra and inter cell-line comparisons among various chromatin regions for four human cell-lines (K562, GM12878, IMR90 and H1hESC), which shows that the openness of chromatin region is well correlated with chromatin function. This method has been executed by an automatic pipeline (AutoChrom3D) and thus can be conveniently used. PMID:23965308

  20. Sub-100 nm biodegradable nanoparticles: in vitro release features and toxicity testing in 2D and 3D cell cultures

    A big challenge in tumor targeting by nanoparticles (NPs), taking advantage of the enhanced permeability and retention effect, is the fabrication of small size devices for enhanced tumor penetration, which is considered fundamental to improve chemotherapy efficacy. The purposes of this study are (i) to engineer the formulation of doxorubicin-loaded poly(d,l-lactic-co-glycolic acid) (PLGA)–block–poly(ethylene glycol) (PEG) NPs to obtain <100 nm devices and (ii) to translate standard 2D cytotoxicity studies to 3D collagen systems in which an initial step gradient of the NPs is present. Doxorubicin release can be prolonged for days to weeks depending on the NP formulation and the pH of the release medium. Sub-100 nm NPs are effectively internalized by HeLa cells in 2D and are less cytotoxic than free doxorubicin. In 3D, <100 nm NPs are significantly more toxic than larger ones towards HeLa cells, and the cell death rate is affected by the contributions of drug release and device transport through collagen. Thus, the reduction of NP size is a fundamental feature from both a technological and a biological point of view and must be properly engineered to optimize the tumor response to the NPs. (paper)

  1. On Dynamic Characteristics of Culture and its Implications

    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.

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

    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...

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

    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. PMID:26215372

  4. RCCS Bioreactor-Based Modelled Microgravity Induces Significant Changes on In Vitro 3D Neuroglial Cell Cultures

    Caterina Morabito

    2015-01-01

    Full Text Available We propose a human-derived neuro-/glial cell three-dimensional in vitro model to investigate the effects of microgravity on cell-cell interactions. A rotary cell-culture system (RCCS bioreactor was used to generate a modelled microgravity environment, and morphofunctional features of glial-like GL15 and neuronal-like SH-SY5Y cells in three-dimensional individual cultures (monotypic aggregates and cocultures (heterotypic aggregates were analysed. Cell survival was maintained within all cell aggregates over 2 weeks of culture. Moreover, compared to cells as traditional static monolayers, cell aggregates cultured under modelled microgravity showed increased expression of specific differentiation markers (e.g., GL15 cells: GFAP, S100B; SH-SY5Y cells: GAP43 and modulation of functional cell-cell interactions (e.g., N-CAM and Cx43 expression and localisation. In conclusion, this culture model opens a wide range of specific investigations at the molecular, biochemical, and morphological levels, and it represents an important tool for in vitro studies into dynamic interactions and responses of nervous system cell components to microgravity environmental conditions.

  5. Type conversion of secretomes in a 3D TAM2 and HCC cell co-culture system and functional importance of CXCL2 in HCC.

    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 cultured with SMMC7721 cells to reconstruct the tumor microenvironment. Conditioned medium from 3D single cultures of M2, SMMC7721 cells, and their co-culture system were analyzed using quantitative proteomics via iTRAQ labeling combined with mass spectrometric analysis. Secretome analysis revealed a total of 159 differential secreted proteins in the co-culture system compared to the single culture systems, with 63 being up-regulated (>1.3-fold) and 96 down-regulated (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

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

    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.

  7. The myofibroblast markers α-SM actin and β-actin are differentially expressed in 2 and 3-D culture models of fibrotic and normal skin.

    Vozenin, M C; Lefaix, J L; Ridi, R; Biard, D S; Daburon, F; Martin, M

    1998-01-01

    To characterize the differences between fibrotic myofibroblasts and normal fibroblasts, we studied two differentiation markers: α-smooth muscle (SM) actin, a specific marker of myofibroblast differentiation, and β-actin, which is overexpressed in the fibrotic tissue. Experiments were performed on fibroblasts isolated from normal pig skin and on subcutaneous myofibroblasts isolated from pig radiation-induced fibrosis. Three culture models were used: cells in monolayers, equivalent dermis, consisting of fibroblasts embedded into a matrix composed of type I collagen, and in vitro reconstituted skin, in which the matrix and containing life fibroblasts were overlaid with keratinocytes. Samples were studied using immunofluorescence and western-blotting. In monolayers cultures, both fibrosis and normal cells expressed α-SM actin. Furthermore, similar amounts of β-actin protein were found. In these conditions, the resulting alterations in the phenotypes of cells made comparison of cultured fibrotic and normal cells irrelevant. Under the two 3-D culture models, normal fibroblasts no longer expressed α-SM actin. They expressed β-actin at the basal level. Moreover, the fibrotic myofibroblasts in both 3-D models retained their differentiation features, expressing α-SM actin and overexpressing β-actin. We found that this normalization was mainly related to the genomic programmation acquired by the cells in the tissue. Cellular motility and microenvironment were also involved, whereas cellular proliferation was not a major factor. Consequently, both three-dimensional models allowed the study of radiation-induced fibrosis in vitro, provided good extrapolations to in vivo conditions and avoided certain of culture artefacts. PMID:22359004

  8. Reduced cytotoxicity and enhanced bioactivity of cationic antimicrobial peptides liposomes in cell cultures and 3D epidermis model against HSV.

    Ron-Doitch, Sapir; Sawodny, Beate; Kühbacher, Andreas; David, Mirjam M Nordling; Samanta, Ayan; Phopase, Jaywant; Burger-Kentischer, Anke; Griffith, May; Golomb, Gershon; Rupp, Steffen

    2016-05-10

    Cationic antimicrobial peptides (AMPs) are part of the innate immunity, and act against a wide variety of pathogenic microorganisms by perturbation of the microorganism's plasma membrane. Although attractive for clinical applications, these agents suffer from limited stability and activity in vivo, as well as non-specific interaction with host biological membranes, leading to cytotoxic adverse effects. We hypothesized that encapsulation of AMPs within liposomes could result in reduced cytotoxicity, and with enhanced stability as well as bioactivity against herpes simplex virus 1 (HSV-1). We formulated nano-sized liposomal formulations of LL-37 and indolicidin, and their physicochemical properties, cellular uptake, in vitro cytotoxicity and antiviral efficacy have been determined. Lower cytotoxicity of LL-37 liposomes was found in comparison to indolicidin liposomes attributed to the superior physicochemical properties, and to the different degree of interaction with the liposomal membrane. The disc-like shaped LL-37 liposomes (106.8±10.1nm, shelf-life stability of >1year) were taken up more rapidly and to a significantly higher extent than the free peptide by human keratinocyte cell line (HaCaT), remained intact within the cells, followed by release of the active peptide within the cytoplasm and migration of the vesicles' lipids to the plasma membrane. LL-37 liposomes were found significantly less toxic than both the free agent and liposomal indolicidin. In the new 3D epidermis model (immortalized primary keratinocytes) liposomal LL-37 treatment (>20μM), but not free LL-37, efficiently protected the epidermis, inhibiting HSV-1 infection. This positive antiviral effect was obtained with no cytotoxicity even at very high concentrations (400μM). Thus, the antiviral activity of encapsulated LL-37 was significantly improved, expanding its therapeutic window. Liposomal LL-37 appears to be a promising delivery system for HSV therapy. PMID:27012977

  9. 3D seismic analysis of the Collyhurst Sandstone: implications for CO2 sequestration in the East Irish Sea Basin

    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

  10. Future Research Challenges for a Computer-Based Interpretative 3D Reconstruction of Cultural Heritage - A German Community's View

    Münster, S.; Kuroczyński, P.; Pfarr-Harfst, M.; Grellert, M.; Lengyel, D.

    2015-08-01

    The workgroup for Digital Reconstruction of the Digital Humanities in the German-speaking area association (Digital Humanities im deutschsprachigen Raum e.V.) was founded in 2014 as cross-disciplinary scientific society dealing with all aspects of digital reconstruction of cultural heritage and currently involves more than 40 German researchers. Moreover, the workgroup is dedicated to synchronise and foster methodological research for these topics. As one preliminary result a memorandum was created to name urgent research challenges and prospects in a condensed way and assemble a research agenda which could propose demands for further research and development activities within the next years. The version presented within this paper was originally created as a contribution to the so-called agenda development process initiated by the German Federal Ministry of Education and Research (BMBF) in 2014 and has been amended during a joint meeting of the digital reconstruction workgroup in November 2014.

  11. Coupling curvature-dependent and shear stress-stimulated neotissue growth in dynamic bioreactor cultures: a 3D computational model of a complete scaffold.

    Guyot, Y; Papantoniou, I; Luyten, F P; Geris, L

    2016-02-01

    The main challenge in tissue engineering consists in understanding and controlling the growth process of in vitro cultured neotissues toward obtaining functional tissues. Computational models can provide crucial information on appropriate bioreactor and scaffold design but also on the bioprocess environment and culture conditions. In this study, the development of a 3D model using the level set method to capture the growth of a microporous neotissue domain in a dynamic culture environment (perfusion bioreactor) was pursued. In our model, neotissue growth velocity was influenced by scaffold geometry as well as by flow- induced shear stresses. The neotissue was modeled as a homogenous porous medium with a given permeability, and the Brinkman equation was used to calculate the flow profile in both neotissue and void space. Neotissue growth was modeled until the scaffold void volume was filled, thus capturing already established experimental observations, in particular the differences between scaffold filling under different flow regimes. This tool is envisaged as a scaffold shape and bioprocess optimization tool with predictive capacities. It will allow controlling fluid flow during long-term culture, whereby neotissue growth alters flow patterns, in order to provide shear stress profiles and magnitudes across the whole scaffold volume influencing, in turn, the neotissue growth. PMID:26758425

  12. Tailored and biodegradable poly(2-oxazoline) microbeads as 3D matrices for stem cell culture in regenerative therapies.

    Lück, Steffen; Schubel, René; Rüb, Jannick; Hahn, Dominik; Mathieu, Evelien; Zimmermann, Heike; Scharnweber, Dieter; Werner, Carsten; Pautot, Sophie; Jordan, Rainer

    2016-02-01

    We present the synthesis of hydrogel microbeads based on telechelic poly(2-oxazoline) (POx) crosslinkers and the methacrylate monomers (HEMA, METAC, SPMA) by inverse emulsion polymerization. While in batch experiments only irregular and ill-defined beads were obtained, the preparation in a microfluidic (MF) device resulted in highly defined hydrogel microbeads. Variation of the MF parameters allowed to control the microbead diameter from 50 to 500 μm. Microbead elasticity could be tuned from 2 to 20 kPa by the POx:monomer composition, the POx chain length, net charge of the hydrogel introduced via the monomer as well as by the organic content of the aqueous phase. The proliferations of human mesenchymal stem cells (hMSCs) on the microbeads were studied. While neutral, hydrophilic POx-PHEMA beads were bioinert, excessive colonization of hMSCs on charged POx-PMETAC and POx-PSPMA was observed. The number of proliferated cells scaled roughly linear with the METAC or SPMA comonomer content. Additional collagen I coating further improved the stem cell proliferation. Finally, a first POx-based system for the preparation of biodegradable hydrogel microcarriers is described and evaluated for stem cell culturing. PMID:26686977

  13. Transcending Cultural Borders: Implications for Science Teaching.

    Jegede, Olugbemiro J.; Aikenhead, Glen S.

    1999-01-01

    Reviews collateral learning theory as a cognitive explanation for how pupils cope with disparate worldviews mediated by transcending cultural borders between their everyday culture and the culture of science. Proposes a new pedagogy in which teachers assume the role of culture broker in the classroom to achieve culturally sensitive curriculum and…

  14. 3D video

    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

  15. 3D Animation Essentials

    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

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

    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

  17. 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

    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.

  18. Real-time measurement of hyperpolarized lactate production and efflux as a biomarker of tumor aggressiveness in an MR compatible 3D cell culture bioreactor.

    Sriram, Renuka; Van Criekinge, Mark; Hansen, Ailin; Wang, Zhen J; Vigneron, Daniel B; Wilson, David M; Keshari, Kayvan R; Kurhanewicz, John

    2015-09-01

    We have developed a 3D cell/tissue culture bioreactor compatible with hyperpolarized (HP) (13)C MR and interrogated HP [1-(13)C]lactate production and efflux in human renal cell carcinoma (RCC) cells. This platform is capable of resolving intracellular and extracellular HP lactate pools, allowing the kinetic measurement of lactate production and efflux in the context of cancer aggressiveness and response to therapy. HP (13)C MR studies were performed on three immortalized human renal cell lines: HK2, a normal renal proximal tubule cell line from which a majority of RCCs arise, UMRC6, a cell line derived from a localized RCC, and UOK262, an aggressive and metastatic RCC. The intra- (Lacin ) and extracellular (Lacex ) HP lactate signals were robustly resolved in dynamic (13)C spectra of the cell lines due to a very small but reproducible chemical shift difference (0.031 ± 0.0005 ppm). Following HP [1-(13)C]pyruvate delivery, the ratio of HP Lacin /Lacex was significantly lower for UOK262 cells compared with both UMRC6 and HK2 cells due to a significant (p culture bioreactor to study not only cellular metabolism but also transport. Additionally, this platform offers a sophisticated way to follow therapeutic interventions and screen novel therapies that target lactate export. PMID:26202449

  19. Characterization of novel tumor stroma markers identified by gene expression profiling of human cancer tissues and 3D co-culture models

    The tumor stroma plays an important role in tumorigenesis. During cancer progression it undergoes changes in architecture, gene expression and secretion of proteolytic enzymes that are essential for the invasive and metastatic phenotype of malignant tumors. Cancer associated fibroblasts (Cafes) represent the major cellular component of the stroma and recent studies demonstrated the prognostic and therapeutic significance of CaF-related molecular signatures. The identification and characterization of genes and signaling pathways involved in the molecular interactions between tumor and stromal cells has been the focus of this study. For that purpose we have used two complementary approaches: the identification of novel tumor stroma targets in human colon cancer samples using whole genome Affymetrix GeneChip analysis and the validation of theses targets in a newly established of 3D co-culture model that mimics the cellular and molecular heterogeneity of human cancers. We have demonstrated increased expression of gene sets related to hypoxia, epithelial-to-mesenchymal transition (EMT) and TGFβ pathway activation in CAFs vs their normal counterparts in both systems. The putative TGFβ target IGFBP7 (insulin-like growth factor binding protein 7) was identified as a tumor stroma marker of epithelial cancers and as a tumor antigen in mesenchyme-derived sarcomas. IGFPB7 was shown to promote anchorage-independent growth in malignant mesenchymal cells and malignant epithelial cells with an EMT-phenotype, whereas a tumor suppressor function was observed in tumor epithelial cells. In summary, we have demonstrated that a number of important signaling pathways involved in cancer progression and metastasis are specifically dysregulated in the tumor stroma both in our in vivo screen and in the in vitro 3D model, illustrating the value of these approaches for the identification and characterization of novel stromal markers. (author)

  20. Queer Cultural Capital: Implications for Education

    Pennell, Summer Melody

    2016-01-01

    This article takes the concept of cultural capital from Yosso's (2005) work and transforms the model for queer communities. While Yosso identified five forms of cultural capital in communities of color (familial, aspirational, navigational, resistant, and linguistic), the author identifies an additional form: transgressive. Queer cultural capital…

  1. Role of Ceacam1 in VEGF induced vasculogenesis of murine embryonic stem cell-derived embryoid bodies in 3D culture

    CEACAM1 (carcinoembryonic antigen-related cell adhesion molecule 1), a type I transmembrane glycoprotein involved in cell-cell adhesion has been shown to act as an angiogenic factor for mouse and human endothelial cells. Based on the ability of CEACAM1 to initiate lumen formation in human mammary epithelial cells grown in 3D culture (Matrigel), we hypothesized that murine CEACAM1 may play a similar role in vasculogenesis. In order to test this hypothesis, murine embryonic stem (ES) cells stimulated with VEGF were differentiated into embryoid bodies (EB) for 8 days (- 8-0 d) and transferred to Matrigel in the presence or absence of anti-CEACAM1 antibody for an additional 12 days (0-12 d). In the absence of anti-CEACAM1 antibody or in the presence of an isotype control antibody, the EB in Matrigel underwent extensive sprouting, generating lengthy vascular structures with well-defined lumina as demonstrated by confocal microscopy, electron microscopy, and immunohistochemical analysis. Both the length and architecture of the vascular tubes were inhibited by anti-CEACAM1 mAb CC1, a mAb that blocks the cell-cell adhesion functions of CEACAM1, thus demonstrating a critical role for this cell-cell adhesion molecule in generating and maintaining vasculogenesis. QRT-PCR analysis of the VEGF treated ES cells grown under conditions that convert them to EB revealed expression of Ceacam1 as early as - 5 to - 3 d reaching a maximum at day 0 at which time EBs were transferred to Matrigel, thereafter levels at first declined and then increased over time. Other markers of vasculogenesis including Pecam1, VE-Cad, and Tie-1 were not detected until day 0 when EBs were transferred to Matrigel followed by a steady increase in levels, indicating later roles in vasculogenesis. In contrast, Tie-2 and Flk-1 (VEGFR2) were detected on day five of EB formation reaching a maximum at day 0 on transfer to Matrigel, similar to Ceacam1, but after which Tie-2 declined over time, while Flk-1 increased

  2. Implications of Japanese Culture for Cultural Construction of Chinese Agricultural Enterprises

    Xu, Ming

    2015-01-01

    The cultural construction in agricultural enterprises in China is not so optimistic especially compared with developed countries. In 2015, 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 s...

  3. A study on application of BacT/ALERT 3D liquid culture technology on tuberculosis drug resistance detection%BacT/ALERT 3D 技术在结核杆菌耐药检测中的应用研究

    黄文忠; 王平平; 吴红照; 周燕珍; 胡真宝

    2014-01-01

    目的:比较 BacT/ALERT 3D 法与传统罗氏(L -J)比例法在耐药结核分枝杆菌检测中的差异,评价BacT/ALERT 3D 快速培养系统在结核分枝杆菌耐药检测中的应用价值。方法对219份固体培养结核杆菌阳性的培养物采用 BacT/ALERT 3D 液体培养技术和 L -J 比例法同时进行药敏试验,并进行比对分析。结果BacT/ALERT 3D 法平均检出时间(8.02±3.85)d,比 L -J 比例法检出时间平均缩短约20 d;BacT/ALERT 3D 法检出耐药菌60株,L -J 比例法检出耐药菌79株,差异无统计学意义(P >0.05);以 L -J 比例法结果为判定标准, BacT/ALERT 3D 法对 INH、RFP、EMB 和 SM 4种抗结核药耐药检测结果符合率分别为95.43%、92.69%、95.43%和92.24%。结论与 L -J 比例法比较,BacT/ALERT 3D 药敏检测方法对结核杆菌的耐药检出结果有较高的一致性,BacT/ALERT 3D 法检出时间明显短于 L -J 比例法。%Objective To evaluate the BacT/ALERT 3D liquid culture technology on the detection of drug resistance of Mycobacterium tuberculosis(MTB)and to compare the difference between this technology and Lowenstein -Jensen (L -J) proportion method.Methods BacT/ALERT 3D liquid culture technology and L -J proportion technology were applied to detect the drug resistance of tuberculosis from the positive cultures of 219 solid culture samples.Results The average detection time of BacT/ALERT 3D method was 8.02 ±3.85 d,which was about 20 days shorter than that of L -J proportion method.60 drug resistance strains were found using BacT/ALERT 3D technology,While 79 drug resistance strains were found using L -J proportion technology.There showed no significant difference (P >0.05).The compliance rate of BacT/ALERT 3D method and L -J proportion method on the anti -tuberculosis drugs INH,RFP,EMB and SMwas 95.43%,92.69%,95.43% and 92.24% respectively.Conclusion BacT/ALERT 3D liquid culture technology could detect drug resistant TB

  4. EUROPEANA AND 3D

    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.

  5. ViHAP3D - Final report

    Scopigno, Roberto

    2005-01-01

    Nearly all of our cultural heritage is inherently three-dimensional. Recent hard- and software developments enabled 3D computer graphics to be one of the most powerful means to represent complex data sets. The ViHAP3D project (ViHAP3D is an acronym for Virtual Heritage - High Quality 3D Acquisition and Presentation) aimed therefore at preserving, presenting, accessing, and promoting cultural heritage using interactive, high-quality 3D graphics. The vision of the project was to create an exact...

  6. Solid works 3D

    This book explains modeling of solid works 3D and application of 3D CAD/CAM. The contents of this book are outline of modeling such as CAD and 2D and 3D, solid works composition, method of sketch, writing measurement fixing, selecting projection, choosing condition of restriction, practice of sketch, making parts, reforming parts, modeling 3D, revising 3D modeling, using pattern function, modeling necessaries, assembling, floor plan, 3D modeling method, practice floor plans for industrial engineer data aided manufacturing, processing of CAD/CAM interface.

  7. Chinese Cultural Implications for ERP Implementation

    Mukesh Srivastava; Betsy Gips

    2009-01-01

    Implementation of an enterprise resource planning (ERP) system in a global environment can be fragmented due to the internal enterprise culture, which is representative of societal culture. In China, this is especially true due to the nationalistic culture of business. The way ERP systems are perceived, treated, and integrated within the business plays a critical role in the success or failure of the implementation. When a Western developed ERP system is implemented in a country where the cul...

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

    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.

  9. A 3D printed microfluidic device for production of functionalized hydrogel microcapsules for culture and differentiation of human Neuronal Stem Cells (hNSC).

    Alessandri, Kevin; Feyeux, Maxime; Gurchenkov, Basile; Delgado, Christophe; Trushko, Anastasiya; Krause, Karl-Heinz; Vignjević, Daniela; Nassoy, Pierre; Roux, Aurélien

    2016-04-26

    We present here a microfluidic device that generates sub-millimetric hollow hydrogel spheres, encapsulating cells and coated internally with a layer of reconstituted extracellular matrix (ECM) of a few microns thick. The spherical capsules, composed of alginate hydrogel, originate from the spontaneous instability of a multi-layered jet formed by co-extrusion using a coaxial flow device. We provide a simple design to manufacture this device using a DLP (digital light processing) 3D printer. Then, we demonstrate how the inner wall of the capsules can be decorated with a continuous ECM layer that is anchored to the alginate gel and mimics the basal membrane of a cellular niche. Finally, we used this approach to encapsulate human Neural Stem Cells (hNSC) derived from human Induced Pluripotent Stem Cells (hIPSC), which were further differentiated into neurons within the capsules with negligible loss of viability. Altogether, we show that these capsules may serve as cell micro-containers compatible with complex cell culture conditions and applications. These developments widen the field of research and biomedical applications of the cell encapsulation technology. PMID:27025278

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

    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.

  11. Cultural Diversity: Implications For Workplace Management

    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.

  12. Open 3D Projects

    Felician ALECU

    2010-01-01

    Full Text Available Many professionals and 3D artists consider Blender as being the best open source solution for 3D computer graphics. The main features are related to modeling, rendering, shading, imaging, compositing, animation, physics and particles and realtime 3D/game creation.

  13. 3d-3d correspondence revisited

    Chung, Hee-Joong; Dimofte, Tudor; Gukov, Sergei; Sułkowski, Piotr

    2016-04-01

    In fivebrane compactifications on 3-manifolds, we point out the importance of all flat connections in the proper definition of the effective 3d {N}=2 theory. The Lagrangians of some theories with the desired properties can be constructed with the help of homological knot invariants that categorify colored Jones polynomials. Higgsing the full 3d theories constructed this way recovers theories found previously by Dimofte-Gaiotto-Gukov. We also consider the cutting and gluing of 3-manifolds along smooth boundaries and the role played by all flat connections in this operation.

  14. IZDELAVA TISKALNIKA 3D

    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 ...

  15. Cultural Diversity: Implications For Workplace Management

    Donatus I. Amaram

    2011-01-01

    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 cul...

  16. 3D and Education

    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?

  17. Mobile 3D tomograph

    Mobile tomographs often have the problem that high spatial resolution is impossible owing to the position or setup of the tomograph. While the tree tomograph developed by Messrs. Isotopenforschung Dr. Sauerwein GmbH worked well in practice, it is no longer used as the spatial resolution and measuring time are insufficient for many modern applications. The paper shows that the mechanical base of the method is sufficient for 3D CT measurements with modern detectors and X-ray tubes. CT measurements with very good statistics take less than 10 min. This means that mobile systems can be used, e.g. in examinations of non-transportable cultural objects or monuments. Enhancement of the spatial resolution of mobile tomographs capable of measuring in any position is made difficult by the fact that the tomograph has moving parts and will therefore have weight shifts. With the aid of tomographies whose spatial resolution is far higher than the mechanical accuracy, a correction method is presented for direct integration of the Feldkamp algorithm

  18. 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

    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

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

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

    2012-01-01

    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 ra...

  20. 3D virtuel udstilling

    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....

  1. Implications of Japanese Culture for Cultural Construction of Chinese Agricultural Enterprises

    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.

  2. Using Cultural Diversity in Teaching Economics: Global Business Implications

    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.…

  3. Underwater 3D filming

    Roberto Rinaldi

    2014-12-01

    Full Text Available After an experimental phase of many years, 3D filming is now effective and successful. Improvements are still possible, but the film industry achieved memorable success on 3D movie’s box offices due to the overall quality of its products. Special environments such as space (“Gravity” and the underwater realm look perfect to be reproduced in 3D. “Filming in space” was possible in “Gravity” using special effects and computer graphic. The underwater realm is still difficult to be handled. Underwater filming in 3D was not that easy and effective as filming in 2D, since not long ago. After almost 3 years of research, a French, Austrian and Italian team realized a perfect tool to film underwater, in 3D, without any constrains. This allows filmmakers to bring the audience deep inside an environment where they most probably will never have the chance to be.

  4. CULTURAL IMPLICATIONS OF A GLOBAL CONTEXT

    Dr. Agnes Erich

    2008-10-01

    Full Text Available At present, we have moved from „a world dominated by cultural isolation in a world where intercultural factors dominate, from an era characterized by cultural autonomy of traditional isolated groups to an era of generalized interrelations and communication”1. Our time has the great historical privilege of moving from a world of isolated civilizations, based to some extent on different spaces and times, to a single world, which is characterized by the same space (world market and the same time (synchronicity of all events. The positive meaning of globalization is that of progress. Accordingly, among many other aspects of globalization, there is an increase in the role of modern information and communication means used in large structures of information and documentation. The major difference between industrial and information age is that in the new economy the information consumers are also its producers, and mediators of information need to be aware who their „customers” are. Particular attention should be paid to the concept of information goods, these being considered the products and services that can be distributed in digital form, such as a book, a movie or a phone conversation.

  5. Earthquake relocation using a 3D a-priori geological velocity model from the western Alps to Corsica: Implication for seismic hazard

    Béthoux, Nicole; Theunissen, Thomas; Beslier, Marie-Odile; Font, Yvonne; Thouvenot, François; Dessa, Jean-Xavier; Simon, Soazig; Courrioux, Gabriel; Guillen, Antonio

    2016-02-01

    The region between the inner zones of the Alps and Corsica juxtaposes an overthickened crust to an oceanic domain, which makes difficult to ascertain the focal depth of seismic events using routine location codes and average 1D velocity models. The aim of this article is to show that, even with a rather lose monitoring network, accurate routine locations can be achieved by using realistic 3D modelling and advanced location techniques. Previous earthquake tomography studies cover the whole region with spatial resolutions of several tens of kilometres on land, but they fail to resolve the marine domain due to the absence of station coverage and sparse seismicity. To overcome these limitations, we first construct a 3D a-priori P and S velocity model integrating known geophysical and geological information. Significant progress has been achieved in the 3D numerical modelling of complex geological structures by the development of dedicated softwares (e.g. 3D GeoModeller), capable at once of elaborating a 3D structural model from geological and geophysical constraints and, possibly, of refining it by inversion processes (Calcagno et al., 2008). Then, we build an arrival-time catalogue of 1500 events recorded from 2000 to 2011. Hypocentres are then located in this model using a numerical code based on the maximum intersection method (Font et al., 2004), updated by Theunissen et al. (2012), as well as another 3D location technique, the NonLinLoc software (Lomax and Curtis, 2001). The reduction of arrival-time residuals and uncertainties (dh, dz) with respect to classical 1D locations demonstrates the improved accuracy allowed by our approach and confirms the coherence of the 3D geological model built and used in this study. Our results are also compared with previous works that benefitted from the installation of dense temporary networks surrounding the studied epicentre area. The resulting 3D location catalogue allows us to improve the regional seismic hazard assessment

  6. Blender 3D cookbook

    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'

  7. 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

    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.

  8. 3D Digital Modelling

    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...

  9. Professional Papervision3D

    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.

  10. Cross-Cultural Discussions in a 3D Virtual Environment and Their Affordances for Learners' Motivation and Foreign Language Discussion Skills

    Jauregi, Kristi; Kuure, Leena; Bastian, Pim; Reinhardt, Dennis; Koivisto, Tuomo

    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 for a period of 5 weeks. The case study had two main objectives: (1) to study the impact that the discussion tasks undertaken in a virtual environment…

  11. Secondary activation of c-abl may be related to translocation to the nucleolar organizer region in an in vitro cultured rat leukemia cell line (K3D)

    Localization of cellular oncogenes (c-onc) near the break points of translocations in tumor cells has indicated involvement of these genes in neoplastic growth. Enhanced transcription of the cellular homolog (c-abl) of the transforming sequence of Abelson murine leukemia virus was observed in K3D, which was one of the cloned cell lines of 7,12-dimethylbenz[a]anthracene-induced rat erythroblastic leukemia. Since the c-abl activation was not observed in the parent cell line (K2D) from which K3D was derived and the latter was different from the former in the presence of a new marker chromosome, t(3;12), this marker may play a role in the expression of c-abl in K3D cells. In contrast to the human c-onc assignments, few rat c-onc assignments have been reported. In situ molecular hybridization studies assigned c-abl to the 3q12 site of the normal chromosome 3 and to the break point of the translocation t(3;12) in K3D cells. Another break point in this translocation chromosome 12p11 involves the nucleolar region, and the 3;12 translocation may involve c-abl and nucleolar cistrons. A cloned v-abl-specific probe was used for hybridization and labelled with 125I-dCTP. These results provide evidence of secondary c-onc activation during karyotypic evolution of cloned malignant cells

  12. Image-Based and Range-Based 3d Modelling of Archaeological Cultural Heritage: the Telamon of the Temple of Olympian ZEUS in Agrigento (italy)

    Lo Brutto, M.; Spera, M. G.

    2011-09-01

    The Temple of Olympian Zeus in Agrigento (Italy) was one of the largest temple and at the same time one of the most original of all the Greek architecture. We don't know exactly how it was because the temple is now almost completely destroyed but it is very well-known for the presence of the Telamons. The Telamons were giant statues (about 8 meters high) probably located outside the temple to fill the interval between the columns. In accordance with the theory most accredited by archaeologists the Telamons were a decorative element and also a support for the structure. However, this hypothesis has never been scientifically proven. One Telamon has been reassembled and is shown at the Archaeological Museum of Agrigento. In 2009 a group of researchers at the University of Palermo has begun a study to test the hypothesis that the Telamons support the weight of the upper part of the temple. The study consists of a 3D survey of the Telamon, to reconstruct a detailed 3D digital model, and of a structural analysis with the Finite Element Method (FEM) to test the possibility that the Telamon could to support the weight of the upper portion of the temple. In this work the authors describe the 3D survey of Telamon carry out with Range-Based Modelling (RBM) and Image-Based Modeling (IBM). The RBM was performed with a TOF laser scanner while the IBM with the ZScan system of Menci Software and Image Master of Topcon. Several tests were conducted to analyze the accuracy of the different 3D models and to evaluate the difference between laser scanning and photogrammetric data. Moreover, an appropriate data reduction to generate a 3D model suitable for FEM analysis was tested.

  13. Use of INSAT-3D sounder and imager radiances in the 4D-VAR data assimilation system and its implications in the analyses and forecasts

    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.

  14. Evaluating the social and cultural implications of the internet

    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...

  15. Non-invasive real-time monitoring by alamarBlue® during in vitro culture of 3D tissue engineered bone constructs

    Zhou, Xiaohua; Holsbeeks, Inge; Impens, Saartje; Sonnaert, Maarten; Bloemen, Veerle; Luyten, Frank Prosper; Schrooten, Jan

    2013-01-01

    Bone Tissue Engineering (TE) aims to develop reproducible and predictive three-dimensional (3D) TE constructs, defined as cell-seeded scaffolds produced by a controlled in vitro process, to heal or replace damaged and non-functional bone. To control and assure the quality of the bone TE constructs, a prerequisite for regulatory authorization, there is a need to develop non-invasive analysis techniques to evaluate TE constructs and to monitor their behavior in real time during in vitro culturi...

  16. 3D Spectroscopic Instrumentation

    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...

  17. 3D Projection Installations

    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....

  18. Herramientas SIG 3D

    Francisco R. Feito Higueruela

    2010-04-01

    Full Text Available Applications of Geographical Information Systems on several Archeology fields have been increasing during the last years. Recent avances in these technologies make possible to work with more realistic 3D models. In this paper we introduce a new paradigm for this system, the GIS Thetrahedron, in which we define the fundamental elements of GIS, in order to provide a better understanding of their capabilities. At the same time the basic 3D characteristics of some comercial and open source software are described, as well as the application to some samples on archeological researchs

  19. Bootstrapping 3D fermions

    Iliesiu, Luca; Kos, Filip; Poland, David; Pufu, Silviu S.; Simmons-Duffin, David; Yacoby, Ran

    2016-03-01

    We study the conformal bootstrap for a 4-point function of fermions in 3D. We first introduce an embedding formalism for 3D spinors and compute the conformal blocks appearing in fermion 4-point functions. Using these results, we find general bounds on the dimensions of operators appearing in the ψ × ψ OPE, and also on the central charge C T . We observe features in our bounds that coincide with scaling dimensions in the GrossNeveu models at large N . We also speculate that other features could coincide with a fermionic CFT containing no relevant scalar operators.

  20. TOWARDS: 3D INTERNET

    Ms. Swapnali R. Ghadge

    2013-01-01

    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 o...

  1. 3D Dental Scanner

    Kotek, L.

    2015-01-01

    This paper is about 3D scan of plaster dental casts. The main aim of the work is a hardware and software proposition of 3D scan system for scanning of dental casts. There were used camera, projector and rotate table for this scanning system. Surface triangulation was used, taking benefits of projections of structured light on object, which is being scanned. The rotate table is controlled by PC. The camera, projector and rotate table are synchronized by PC. Controlling of stepper motor is prov...

  2. Interaktiv 3D design

    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....

  3. 3D Harmonic Echocardiography:

    M.M. Voormolen

    2007-01-01

    textabstractThree dimensional (3D) echocardiography has recently developed from an experimental technique in the ’90 towards an imaging modality for the daily clinical practice. This dissertation describes the considerations, implementation, validation and clinical application of a unique

  4. Tangible 3D Modelling

    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...

  5. Shaping 3-D boxes

    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...

  6. 3D structure prediction of human β1-adrenergic receptor via threading-based homology modeling for implications in structure-based drug designing.

    Zaheer Ul-Haq

    Full Text Available Dilated cardiomyopathy is a disease of left ventricular dysfunction accompanied by impairment of the β1-adrenergic receptor (β1-AR signal cascade. The disturbed β1-AR function may be based on an elevated sympathetic tone observed in patients with heart failure. Prolonged adrenergic stimulation may induce metabolic and electrophysiological disturbances in the myocardium, resulting in tachyarrhythmia that leads to the development of heart failure in human and sudden death. Hence, β1-AR is considered as a promising drug target but attempts to develop effective and specific drug against this tempting pharmaceutical target is slowed down due to the lack of 3D structure of Homo sapiens β1-AR (hsβADR1. This study encompasses elucidation of 3D structural and physicochemical properties of hsβADR1 via threading-based homology modeling. Furthermore, the docking performance of several docking programs including Surflex-Dock, FRED, and GOLD were validated by re-docking and cross-docking experiments. GOLD and Surflex-Dock performed best in re-docking and cross docking experiments, respectively. Consequently, Surflex-Dock was used to predict the binding modes of four hsβADR1 agonists. This study provides clear understanding of hsβADR1 structure and its binding mechanism, thus help in providing the remedial solutions of cardiovascular, effective treatment of asthma and other diseases caused by malfunctioning of the target protein.

  7. TOOTEKO: A CASE STUDY OF AUGMENTED REALITY FOR AN ACCESSIBLE CULTURAL HERITAGE. DIGITIZATION, 3D PRINTING AND SENSORS FOR AN AUDIO-TACTILE EXPERIENCE

    D’Agnano, F.; C. Balletti; Guerra, F.; Vernier, P

    2015-01-01

    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...

  8. Tooteko: a Case Study of Augmented Reality for AN Accessible Cultural Heritage. Digitization, 3d Printing and Sensors for AN Audio-Tactile Experience

    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

  9. From Mental Game to Cultural Praxis: A Cultural Studies Model's Implications for the Future of Sport Psychology

    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…

  10. Experimental Study on Self-assembly of KLD-12 Peptide Hydrogel and 3-D Culture of MSC Encapsulated within Hydrogel In Vitro

    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.