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3D CELL CULTURE

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Created: 09 Sep 2015

Resource/NewsletterV4

Volume 4, June 2016

Newsletter

3D at Purdue

3D in Focus

3D in Publications

3D in Meetings

3D at Purdue

Standardized, Self-assembling Type I Collagens for Modernized In-vitro 3D Human Tumor-Stroma Models — Shooting for the Moon

By: Catherine F. Whittington, PHD LIFA Postdoctoral Fellow

The Harbin Laboratory has a rich history in the study of interactions between the cell and its surrounding 3D extracellular matrix (ECM). Early on, the laboratory recognized that traditional 2D in vitro models for many events of development and disease were not amenable to systematic modulation or mechanistic study of specific cell-ECM interactions. Therefore, we developed an uncommon set of type I collagen building blocks (oligomers)^{1, 3} which retain their inherent fibril-forming capacity and can be used to predictably and reproducibly control fibril- and matrix-level properties of polymerizable matrices to recapitulate what is found in the body’s tissues.^{6, 10}

We have shown that these 3D collagen matrices have biochemical and biophysical bioinstructive function and can be tuned to predictably guide lineage-specific differentiation of mesenchymal stem cells into bone for next generation tissue engineered products or direct vessel network formation by human endothelial colony forming cells (hECFC) for therapeutic vascularization strategies (Figure 1).^{2, 4, 7, 8, 11}

We also use our type I collagen oligomer formulations as a tool for more basic biological research to actively study cell-ECM interactions that are important for drug discovery, toxicity testing, and a myriad of biological activities including disease development and progression.^{5, 9}

A current collaboration with Eli Lilly and Company in Indianapolis, sponsored by the Lilly Innovation Fellowship Award Program, allows us to adapt our collagen technology within a pharmaceutic industry context as a platform to improve upon preclinical tumor models that typically lack the ability to reliably reproduce complex biological signaling inherent to the tumor microenvironment in vivo. Our work seeks to tackle Eli Lilly’s grand challenge of “establishing clinical efficacy and safety earlier in the drug development process” by addressing the limitations of current tumor models (e.g., 2D cell culture and spheroids) through the incorporation of 3D ECMs into existing target validation and drug sensitivity testing methods. At the same time, we are also working to improve how we evaluate efficacy in 3D by adapting existing analysis techniques and high throughput systems to accommodate 3D cultures.

Collectively, this effort is working to translate new tools and a better understanding of the role of the tumor microenvironment in cancer initiation, progression, metastasis, and therapeutic responsiveness, a goal consistent with the National Cancer MoonShot Initiative.

Collectively, this effort is working to translate new tools and a better understanding of the role of the tumor microenvironment in cancer initiation, progression, metastasis, and therapeutic responsiveness, a goal consistent with the National Cancer MoonShot Initiative.

Figure 1: Schematic representation of a 3D in-vitro vasculogenesis model in which hECFC were suspended within type I collagen to form a vascularized tissue construct. Oligomer collagen matrices supported vascular networks (Green) with lumen that were further stabilized via basement membrane deposition (Red; Bottom image). (Whittington, C.F., Yoder, M.C., Voytik-Harbin, S.L. Macromolecular Bioscience, 13:1135-1149, 2013. )

Recent work from the laboratory focused on multiple cancer types, including colorectal, pancreatic, breast⁵, and brain⁹ cancers, used 3D collagen-based ECMs to study how ECM composition and biophysical properties modulate the epithelial to mesenchymal transition (EMT) of cancer cells, which plays a significant role in tumor metastasis and drug resistance. In our study, we used our type I collagen oligomers to study the mechanisms of EMT mechanobiology and found that ECM composition and biophysical properties (stiffness) are important for guiding EMT (Figure 2). Cells not only behaved differently in terms of their EMT status when exposed to a basement membrane (BM; sheet-like; Matrigel) ECM compared to an interstitial (IM; fibrillar; type I collagen) ECM, but they also exhibited varied drug sensitivity. These results provide new insight into how IM fibril microstructure and mechanical properties guide EMT. They also challenge existing correlations between stromal properties and EMT established using conventional preclinical models, and help identify critical parameters for engineering pathophysiologically relevant tumor-stroma models.

As we continue to use 3D ECMs in our studies, we gain more insight into how they may contribute to the development of new therapeutic strategies. With each new application we pursue, discoveries are made that reinforce the importance of the 3D ECM in regulating cell behavior in development and disease. As such, we advocate for researchers to consider the ECM as a key component necessary to produce more physiologically relevant models that can help us accelerate the transition of laboratory innovations from bench to bedside.

Figure 2: (A) Schematic representation of EMT progression as epithelial cells (rounded, grouped) break free of the basement membrane (BM) and become more mesenchymal (spindle-shaped) to invade the interstitial matrix (IM). (B) HT29 colorectal cancer cells (Green=F-actin; Blue=nucleus) undergo morphological changes associated with EMT when exposed to decreasing ratios of Matrigel (representing BM) to type I collagen oligomer (representing IM; White=collagen fibrils). Scale bar = 50 µm.

Project Contributors:

LIFA Postdoctoral Fellow: Catherine F. Whittington
PHD Purdue Faculty Mentor: Sherry L. Harbin, PHD
Eli Lilly Oncology Group Mentors: Shripad Bhagwat, PHD and Sheng-Bin Peng, PHD
Purdue Biomedical Engineering Graduate Student: T.J. Puls
Purdue Biomedical Engineering Undergraduate Student: Xiaohong Tan

Selected Relevant Publications:

Kreger S.T., Bell, B.J., Bailey, J., Stites, E., Kuske, J., Waisner, B., and Voytik-Harbin, S.L. Polymerization and matrix physical properties as important design considerations for soluble collagen formulations. Biopolymers, 93:690-707, 2010.
Critser, P.J., Kreger, S.T., Voytik-Harbin, S.L., and Yoder, M.C. Collagen matrix physical properties modulate endothelial colony forming cell derived vessels in vivo. Microvasc Res 80:23-30, 2010.
Bailey, J.L., Critser, P.J., Whittington, C., Kuske, J.L., Yoder, M.C., and Voytik-Harbin, S.L. Collagen oligomers modulate physical and biological properties of three-dimensional self-assembled matrices. Biopolymers 95:77-93, 2011.
Critser, P.J., Voytik-Harbin, S.L., and Yoder, M.C. Isolating and defining cells to engineer human blood vessels. Cell Prolif 44:15-21, 2011.
Monteleon, C.L., Hartsell, A., Sedgwick, A., Whittington, C., Voytik-Harbin, S., and D’Souza- Schorey, C. Inverted Epithelial Cysts: Interlinked roles for ARF6, Rac1 and the matrix microenvironment. Mol Biol Cell 23:4495-505, 2012
Whittington, C., Brandner, E., Teo, K.Y., Han, B., Nauman, E., and Voytik-Harbin, S.L. Oligomers modulate interfibril branching and mass transport properties of collagen matrices. Microsc Microanal 19:1323-1333, 2013.
Kim, S.J., Wan, Q., Cho, E., Han, B., Yoder, M.C., Voytik-Harbin, S.L., and Na., S. Matrix rigidity regulates spatiotemporal dynamics of Cdc42 activity and vacuole formation kinetics of endothelial colony forming cells. Biochem Biophys Res Commun 443:1280-1285, 2014.
Richardson, M.R., Robbins, E.P., Vemula, S., Critser, P.J., Whittington, C., Voytik-Harbin, S.L., and Yoder, M.C. Angiopoietin-like protein 2 regulates endothelial colony forming cell vasculogenesis. Angiogenesis 17:675-683, 2014.
Herrera-Perez M, Voytik-Harbin SL, Rickus JL. Extracellular matrix properties regulate the migratory response of glioblastoma stem cells in three-dimensional culture. Tissue Eng Part A 21:2572-82, 2015.
Blum, K.M., Novak, T., Watkins, L., Neu, C.P., Wallace, J.M., Bart, Z.R. and Voytik-Harbin, S.L. Acellular and cellular high-density, collagen-fibril constructs with suprafibrillar organization. Biomater Sci 4:711-723, 2016.
Buno, K.P., Chen, X., Weibel, J.A., Thiede, S.N., Garimella, S.V., Yoder, M.C., and Voytik- Harbin, S.L. In Vitro Multitissue Interface model supports rapid vasculogenesis and mechanistic study of vascularization across tissue compartments. ACS Appl Mater Interfaces 2016 (Epub ahead of print).

3D in Focus

Within the body, cells reside in tissues with distinct physiological microenvironments or niches. Increasing evidence illustrates that the adherent cells cultured as a two-dimensional (2D) monolayer on a plastic surface lack the type of interactions with the microenvironment that normally occur in vivo. The cells cultured under three-dimensional (3D) conditions are enticed to assemble into structures with aspects of the physiologically relevant organization and function of specific tissues. In fact, the more specialized the niche created thanks to appropriate 3D culture conditions, the more exquisite the regulation of cell behavior including cell proliferation, migration, differentiation, survival, etc. For further information on this topic please refer to our discussion below of the article by Marinkovic et al.

Marinkovic M, Block TJ, Rakian R, Li Q, Wang E, Reilly MA, Dean DD, Chen XD “One size does not fit all: developing a cell-specific niche for in vitro study of cell behavior” Matrix Biol. (2016) 54–55, 426–441

To address the importance of creating a tissue specific niche in 3D cell culture, the authors prepared an extracellular matrix (ECM) by ‘decellularization’ of microenvironments containing either a matrix synthesized by bone marrow cells (BM-ECM) or a matrix synthesized by adipocytes (AD-ECM). Compared to polystyrene surfaces used for 2D cell culture, the proliferation of mesenchymal stem cells (MSCs) obtained either from the bone marrow or from the adipose tissue was significantly increased (by 1.4 to 2 fold) when cultured in the presence of ECM. The resulting effects depended on the ECM niche (i.e., BM-ECM promoted the proliferation of BM-MSCs over AD-MSCs, and vice versa). Moreover, BM- and AD-ECM preferentially directed the differentiation of both groups of MSCs towards osteogenic lineage and adipogenic lineage, respectively, suggesting a tissue-specific effect of the ECM on MSCs differentiation.

To dissect the characteristics of the tissue specific niches, the chemical (protein composition) and physical (topographical and mechanical) properties of the two ECMs were compared. By mass spectrometry, BM- and AD-ECM were found to share approximately 64% of their protein components, mostly collagen type VI, followed by types XII and I. The compositional variation was the greatest with proteoglycans and glycoproteins.

Both BM- and AD-ECMs exhibited unique topographical and mechanical properties known to regulate stem cell differentiation. A combination of atomic force microscopy, second-harmonic generation microscopy and immunofluorescence microscopy revealed that BM- and AD-ECMs exhibited unique architectures, largely attributable to variations in collagen VI organization. In immunofluorescence microscopy, collagen VI in BM-ECM was characterized by fine, highly aligned fibrils, while AD-ECM contained fibrils that were less ordered and were organized into denser bundles. Moreover, mechanical characterization with small angle oscillatory shear rheology indicated that BM-ECM was three orders of magnitude stiffer than AD-ECM. According to the authors, the unique topographical and mechanical properties of BM- and AD-ECM may be associated with the preferential effect of each ECM on the differentiation of MSCs.

In summary, these experiments conducted with native ECMs (BM-ECM and AD-ECM) that represent the tissue-specific microenvironment (niche) for bone marrow and adipose tissues, outline the importance of developing more sophisticated 3D culture systems in order to replicate the in vivo niches. Such a cell culture goal will require understanding the exact role of individual chemical and physical properties (and/or the combination of these properties) of the ECMs in cell differentiation.

3D in Publications

Complete List -- 3D culture related articles on PubMed (last 2 months)

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3D in Publications

Complete List

Abbott, R. D., Wang, R. Y., Reagan, M. R., Chen, Y., Borowsky, F. E., Zieba, A., Marra, K. G., Rubin, J. P., Ghobrial, I. M., and Kaplan, D. L. (2016). The Use of Silk as a Scaffold for Mature, Sustainable Unilocular Adipose 3D Tissue Engineered Systems. Adv Healthc Mater.

Ahmed, M. I., Elias, S., Mould, A. W., Bikoff, E. K., and Robertson, E. J. (2016). The transcriptional repressor Blimp1 is expressed in rare luminal progenitors and is essential for mammary gland development. Development 143, 1663-1673.

Akasov, R., Zaytseva-Zotova, D., Burov, S., Leko, M., Dontenwill, M., Chiper, M., Vandamme, T., and Markvicheva, E. (2016). Formation of multicellular tumor spheroids induced by cyclic RGD-peptides and use for anticancer drug testing in vitro. Int J Pharm 506, 148-157.

Aleksandrova, A. V., Pulkova, N. P., Gerasimenko, T. N., Anisimov, N. Y., Tonevitskaya, S. A., and Sakharov, D. A. (2016). Mathematical and Experimental Model of Oxygen Diffusion for HepaRG Cell Spheroids. Bull Exp Biol Med 160, 857-860.

Alessandri, K., Feyeux, M., Gurchenkov, B., Delgado, C., Trushko, A., Krause, K. H., Vignjevic, D., Nassoy, P., and Roux, A. (2016). A 3D printed microfluidic device for production of functionalized hydrogel microcapsules for culture and differentiation of human Neuronal Stem Cells (hNSC). Lab Chip 16, 1593-1604.

Alinezhad, S., Vaananen, R. M., Mattsson, J., Li, Y., Tallgren, T., Tong Ochoa, N., Bjartell, A., Akerfelt, M., Taimen, P., Bostrom, P. J., et al. (2016). Validation of Novel Biomarkers for Prostate Cancer Progression by the Combination of Bioinformatics, Clinical and Functional Studies. PLoS One 11, e0155901.

Alonso-Nocelo, M., Abellan-Pose, R., Vidal, A., Abal, M., Csaba, N., Alonso, M. J., Lopez-Lopez, R., and de la Fuente, M. (2016). Selective interaction of PEGylated polyglutamic acid nanocapsules with cancer cells in a 3D model of a metastatic lymph node. J Nanobiotechnology 14, 51.

Anzai, K., Chikada, H., Tsuruya, K., Ida, K., Kagawa, T., Inagaki, Y., Mine, T., and Kamiya, A. (2016). Foetal hepatic progenitor cells assume a cholangiocytic cell phenotype during two-dimensional pre-culture. Sci Rep 6, 28283.

Aoki, T., Yamamoto, K., Fukuda, M., Shimogonya, Y., Fukuda, S., and Narumiya, S. (2016). Sustained expression of MCP-1 by low wall shear stress loading concomitant with turbulent flow on endothelial cells of intracranial aneurysm. Acta Neuropathol Commun 4, 48.

Arrigoni, C., Bongio, M., Talo, G., Bersini, S., Enomoto, J., Fukuda, J., and Moretti, M. (2016). Rational Design of Prevascularized Large 3D Tissue Constructs Using Computational Simulations and Biofabrication of Geometrically Controlled Microvessels. Adv Healthc Mater.

Asthana, A., and Kisaalita, W. S. (2016). Molecular basis for cytokine biomarkers of complex 3D microtissue physiology in vitro. Drug Discov Today 21, 950-961.

Balasubramanian, S., Packard, J. A., Leach, J. B., and Powell, E. M. (2016). Three-dimensional environment sustains morphological heterogeneity and promotes phenotypic progression during astrocyte development. Tissue Eng Part A.

Bandula, S., Magdeldin, T., Stevens, N., Yeung, J., Moon, J. C., Taylor, S. A., Cheema, U., and Punwani, S. (2016). Initial validation of equilibrium contrast imaging for extracellular volume quantification using a three-dimensional engineered tissue model. J Magn Reson Imaging 43, 1224-1229.

Bao, J., Wu, Q., Wang, Y., Li, Y., Li, L., Chen, F., Wu, X., Xie, M., and Bu, H. (2016). Enhanced hepatic differentiation of rat bone marrow-derived mesenchymal stem cells in spheroidal aggregate culture on a decellularized liver scaffold. Int J Mol Med.

Barbier, M., Jaensch, S., Cornelissen, F., Vidic, S., Gjerde, K., de Hoogt, R., Graeser, R., Gustin, E., and Chong, Y. T. (2016). Ellipsoid Segmentation Model for Analyzing Light-Attenuated 3D Confocal Image Stacks of Fluorescent Multi-Cellular Spheroids. PLoS One 11, e0156942.

Basso, F. G., Soares, D. G., de Souza Costa, C. A., and Hebling, J. (2016). Low-level laser therapy in 3D cell culture model using gingival fibroblasts. Lasers Med Sci.

Beaton, H., Andrews, D., Parsons, M., Murphy, M., Gaffney, A., Kavanagh, D., McKay, G. J., Maxwell, A. P., Taylor, C. T., Cummins, E. P., et al. (2016). Wnt6 regulates epithelial cell differentiation and is dysregulated in renal fibrosis. Am J Physiol Renal Physiol, ajprenal.00136.02016.

Bell, C. C., Hendriks, D. F., Moro, S. M., Ellis, E., Walsh, J., Renblom, A., Fredriksson Puigvert, L., Dankers, A. C., Jacobs, F., Snoeys, J., et al. (2016). Characterization of primary human hepatocyte spheroids as a model system for drug-induced liver injury, liver function and disease. Sci Rep 6, 25187.

Blackmon, R. L., Sandhu, R., Chapman, B. S., Casbas-Hernandez, P., Tracy, J. B., Troester, M. A., and Oldenburg, A. L. (2016). Imaging Extracellular Matrix Remodeling In Vitro by Diffusion-Sensitive Optical Coherence Tomography. Biophys J 110, 1858-1868.

Bonomi, A., Steimberg, N., Benetti, A., Berenzi, A., Alessandri, G., Pascucci, L., Boniotti, J., Cocce, V., Sordi, V., Pessina, A., and Mazzoleni, G. (2016). Paclitaxel-releasing mesenchymal stromal cells inhibit the growth of multiple myeloma cells in a dynamic 3D culture system. Hematol Oncol.

Bourget, J. M., Laterreur, V., Gauvin, R., Guillemette, M. D., Miville-Godin, C., Mounier, M., Tondreau, M. Y., Tremblay, C., Labbe, R., Ruel, J., et al. (2016). Microstructured human fibroblast-derived extracellular matrix scaffold for vascular media fabrication. J Tissue Eng Regen Med.

Boye, K., Jacob, H., Frikstad, K. M., Nesland, J. M., Maelandsmo, G. M., Dahl, O., Nesbakken, A., and Flatmark, K. (2016). Prognostic significance of S100A4 expression in stage II and III colorectal cancer: results from a population-based series and a randomized phase III study on adjuvant chemotherapy. Cancer Med.

Branco da Cunha, C., Klumpers, D. D., Koshy, S. T., Weaver, J. C., Chaudhuri, O., Seruca, R., Carneiro, F., Granja, P. L., and Mooney, D. J. (2016). CD44 alternative splicing in gastric cancer cells is regulated by culture dimensionality and matrix stiffness. Biomaterials 98, 152-162.

Brandenberg, N., and Lutolf, M. P. (2016). In Situ Patterning of Microfluidic Networks in 3D Cell-Laden Hydrogels. Adv Mater.

Breslin, S., and O'Driscoll, L. (2016). The relevance of using 3D cell cultures, in addition to 2D monolayer cultures, when evaluating breast cancer drug sensitivity and resistance. Oncotarget.

Casey, A., Gargotti, M., Bonnier, F., and Byrne, H. J. (2016). Chemotherapeutic efficiency of drugs in vitro: Comparison of doxorubicin exposure in 3D and 2D culture matrices. Toxicol In Vitro 33, 99-104.

Castro, N. J., Tan, W. N., Shen, C., and Zhang, L. G. (2016). Simulated Body Fluid Nucleation of 3D Printed Elastomeric Scaffolds for Enhanced Osteogenesis. Tissue Eng Part A.

Celik, E., Bayram, C., Akcapinar, R., Turk, M., and Denkbas, E. B. (2016). The effect of calcium chloride concentration on alginate/Fmoc-diphenylalanine hydrogel networks. Mater Sci Eng C Mater Biol Appl 66, 221-229.

Chen, K., Wu, M., Guo, F., Li, P., Chan, C. Y., Mao, Z., Li, S., Ren, L., Zhang, R., and Huang, T. J. (2016a). Rapid formation of size-controllable multicellular spheroids via 3D acoustic tweezers. Lab Chip.

Chen, W., Ma, J., Zhu, L., Morsi, Y., Ei-Hamshary, H., Al-Deyab, S. S., and Mo, X. (2016b). Superelastic, superabsorbent and 3D nanofiber-assembled scaffold for tissue engineering. Colloids Surf B Biointerfaces 142, 165-172.

Chen, X., and Thibeault, S. L. (2016). Cell-cell interaction between vocal fold fibroblasts and bone marrow mesenchymal stromal cells in three-dimensional hyaluronan hydrogel. J Tissue Eng Regen Med 10, 437-446.

Chi, C. W., Ahmed, A. R., Dereli-Korkut, Z., and Wang, S. (2016). Microfluidic cell chips for high-throughput drug screening. Bioanalysis 8, 921-937.

Chitrangi, S., Nair, P., and Khanna, A. (2016). Three-dimensional polymer scaffolds for enhanced differentiation of human mesenchymal stem cells to hepatocyte-like cells: a comparative study. J Tissue Eng Regen Med.

Cho, I. S., Cho, M. O., Li, Z., Nurunnabi, M., Park, S. Y., Kang, S. W., and Huh, K. M. (2016). Synthesis and characterization of a new photo-crosslinkable glycol chitosan thermogel for biomedical applications. Carbohydr Polym 144, 59-67.

Choi, C. H., Wang, H., Lee, H., Kim, J. H., Zhang, L., Mao, A., Mooney, D. J., and Weitz, D. A. (2016). One-step generation of cell-laden microgels using double emulsion drops with a sacrificial ultra-thin oil shell. Lab Chip 16, 1549-1555.

Clevers, H. (2016). Modeling Development and Disease with Organoids. Cell 165, 1586-1597.

Coleman, C. N., Higgins, G. S., Brown, J. M., Baumann, M., Kirsch, D. G., Willers, H., Prasanna, P. G., Dewhirst, M. W., Bernhard, E. J., and Ahmed, M. M. (2016). Improving the Predictive Value of Preclinical Studies in Support of Radiotherapy Clinical Trials. Clin Cancer Res.

Coloff, J. L., Murphy, J. P., Braun, C. R., Harris, I. S., Shelton, L. M., Kami, K., Gygi, S. P., Selfors, L. M., and Brugge, J. S. (2016). Differential Glutamate Metabolism in Proliferating and Quiescent Mammary Epithelial Cells. Cell Metab 23, 867-880.

Cui, C., Ye, L., Huang, Z., Huang, S., Liu, H., and Yu, J. (2016). FAM172A is a tumor suppressor in colorectal carcinoma. Tumour Biol 37, 6501-6510.

Dai, G., Wan, W., Zhao, Y., Wang, Z., Li, W., Shi, P., and Shen, Y. (2016). Controllable 3D alginate hydrogel patterning via visible-light induced electrodeposition. Biofabrication 8, 025004.

Deharde, D., Schneider, C., Hiller, T., Fischer, N., Kegel, V., Lubberstedt, M., Freyer, N., Hengstler, J. G., Andersson, T. B., Seehofer, D., et al. (2016). Bile canaliculi formation and biliary transport in 3D sandwich-cultured hepatocytes in dependence of the extracellular matrix composition. Arch Toxicol.

Duan, B., Yin, Z., Hockaday Kang, L., Magin, R. L., and Butcher, J. T. (2016). Active tissue stiffness modulation controls valve interstitial cell phenotype and osteogenic potential in 3D culture. Acta Biomater 36, 42-54.

Duarte Campos, D. F., Blaeser, A., Buellesbach, K., Sen, K. S., Xun, W., Tillmann, W., and Fischer, H. (2016). Bioprinting Organotypic Hydrogels with Improved Mesenchymal Stem Cell Remodeling and Mineralization Properties for Bone Tissue Engineering. Adv Healthc Mater 5, 1336-1345.

Eberle, F., Saulich, M. F., Leinberger, F. H., Seeger, W., Engenhart-Cabillic, R., Dikomey, E., Hanze, J., Hattar, K., and Subtil, F. S. (2016). Cancer cell motility is affected through 3D cell culturing and SCF/c-Kit pathway but not by X-irradiation. Radiother Oncol.

Edmondson, R., Adcock, A. F., and Yang, L. (2016). Influence of Matrices on 3D-Cultured Prostate Cancer Cells' Drug Response and Expression of Drug-Action Associated Proteins. PLoS One 11, e0158116.

Emura, M., and Aufderheide, M. (2016). Challenge for 3D culture technology: Application in carcinogenesis studies with human airway epithelial cells. Exp Toxicol Pathol 68, 255-261.

Esch, M. B., Ueno, H., Applegate, D. R., and Shuler, M. L. (2016). Modular, pumpless body-on-a-chip platform for the co-culture of GI tract epithelium and 3D primary liver tissue. Lab Chip.

Fan, Y., Nguyen, D. T., Akay, Y., Xu, F., and Akay, M. (2016). Engineering a Brain Cancer Chip for High-throughput Drug Screening. Sci Rep 6, 25062.

Franco-Barraza, J., Beacham, D. A., Amatangelo, M. D., and Cukierman, E. (2016). Preparation of Extracellular Matrices Produced by Cultured and Primary Fibroblasts. Curr Protoc Cell Biol 71, 10.19.11-10.19.34.

Fu, F., Shang, L., Zheng, F., Chen, Z., Wang, H., Wang, J., Gu, Z., and Zhao, Y. (2016). Cells cultured on core-shell photonic crystal barcodes for drug screening. ACS Appl Mater Interfaces.

Gagliano, N., Celesti, G., Tacchini, L., Pluchino, S., Sforza, C., Rasile, M., Valerio, V., Laghi, L., Conte, V., and Procacci, P. (2016). Epithelial-to-mesenchymal transition in pancreatic ductal adenocarcinoma: Characterization in a 3D-cell culture model. World J Gastroenterol 22, 4466-4483.

Gangadhara, S., Smith, C., Barrett-Lee, P., and Hiscox, S. (2016). 3D culture of Her2+ breast cancer cells promotes AKT to MAPK switching and a loss of therapeutic response. BMC Cancer 16, 345.

Gao, G., Hubbell, K., and Cui, X. (2016). NR2F2 regulates chondrogenesis of human mesenchymal stem cells in bioprinted cartilage. Biotechnol Bioeng.

Garcia-Caballero, M., Blacher, S., Paupert, J., Quesada, A. R., Medina, M. A., and Noel, A. (2016). Novel application assigned to toluquinol: inhibition of lymphangiogenesis by interfering with VEGF-C/VEGFR-3 signalling pathway. Br J Pharmacol 173, 1966-1987.

Gawri, R., Shiba, T., Pilliar, R., and Kandel, R. (2016). Inorganic polyphosphates enhances nucleus pulposus tissue formation in vitro. J Orthop Res.

Gerges, I., Tamplenizza, M., Rossi, E., Tocchio, A., Martello, F., Recordati, C., Kumar, D., Forsyth, N. R., Liu, Y., and Lenardi, C. (2016). A Tailor-Made Synthetic Polymer for Cell Encapsulation: Design Rationale, Synthesis, Chemical-Physics and Biological Characterizations. Macromol Biosci 16, 870-881.

Gilmour, A. D., Woolley, A. J., Poole-Warren, L. A., Thomson, C. E., and Green, R. A. (2016). A critical review of cell culture strategies for modelling intracortical brain implant material reactions. Biomaterials 91, 23-43.

Goliwas, K. F., Miller, L. M., Marshall, L. E., Berry, J. L., and Frost, A. R. (2016). Preparation and Analysis of In Vitro Three Dimensional Breast Carcinoma Surrogates. J Vis Exp.

Gomes, L. R., Vessoni, A. T., and Menck, C. F. (2016). Microenvironment and autophagy cross-talk: Implications in cancer therapy. Pharmacol Res 107, 300-307.

Goppert, B., Sollich, T., Abaffy, P., Cecilia, A., Heckmann, J., Neeb, A., Backer, A., Baumbach, T., Gruhl, F. J., and Cato, A. C. (2016). Superporous Poly(ethylene glycol) Diacrylate Cryogel with a Defined Elastic Modulus for Prostate Cancer Cell Research. Small.

Hagiwara, M., Kawahara, T., and Nobata, R. (2016). Tissue in Cube: In Vitro 3D Culturing Platform with Hybrid Gel Cubes for Multidirectional Observations. Adv Healthc Mater.

Ham, S. L., Joshi, R., Thakuri, P. S., and Tavana, H. (2016). Liquid-based three-dimensional tumor models for cancer research and drug discovery. Exp Biol Med (Maywood) 241, 939-954.

Hamdi, D. H., Chevalier, F., Groetz, J. E., Durantel, F., Thuret, J. Y., Mann, C., and Saintigny, Y. (2016). Comparable Senescence Induction in Three-dimensional Human Cartilage Model by Exposure to Therapeutic Doses of X-rays or C-ions. Int J Radiat Oncol Biol Phys 95, 139-146.

Hammond, T., Allen, P., and Birdsall, H. (2016). Is There a Space-Based Technology Solution to Problems with Preclinical Drug Toxicity Testing? Pharm Res.

Han, H. W., and Hsu, S. H. (2016). Chitosan-hyaluronan based 3D co-culture platform for studying the crosstalk of lung cancer cells and mesenchymal stem cells. Acta Biomater.

Haqq, J., Howells, L. M., Garcea, G., and Dennison, A. R. (2016). Targeting pancreatic cancer using a combination of gemcitabine with the omega-3 polyunsaturated fatty acid emulsion, Lipidem. Mol Nutr Food Res 60, 1437-1447.

Harrison, R., Markides, H., Morris, R. H., Richards, P., El Haj, A. J., and Sottile, V. (2016). Autonomous magnetic labelling of functional mesenchymal stem cells for improved traceability and spatial control in cell therapy applications. J Tissue Eng Regen Med.

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