What is 3D Culture? - part 2

Three-dimensional cultures are in vitro cell culture models in which extracellular matrix (ECM) proteins recapitulate the physiological cell environment to promote cell differentiation and tissue architecture as seen in vivo. These cultures exhibit considerable similarities in their gene expression profiles; morphology; cell–cell and cell–matrix interactions; and differentiation to their originating tissues, while there are significant differences for cells grown on flat 2-D plastic substrata. (1–3) Under 3D culture conditions, epithelial cells organize to resemble their subsequent tissues, (4–6) endothelial cells arrange to form vascular networks, (7) stromal cells maintain a mesenchymal phenotype, (8) and tumor cells assemble into disorganized masses.(9) In these models, the extracellular environment dictates the cellular program and subsequent differentiation and structure formation; thus it is important to select the right ECM proteins based on cell type and function. Cells of epithelial and endothelial origin have an underlying basement membrane orchestrating their growth and development, and this microenvironment is most commonly reconstructed using a basement membrane extract (BME). Alternatively, mesenchymal cells reside in collagen rich connective tissues, and purified collagen I is most commonly used to mimic these surroundings. Other soluble factors may also be necessary to direct cellular differentiation; these include serum, growth factors, hormones, and other cytokines. Most 3D models have optimized conditions to promote proper differentiation, and these parameters can often be found in the scientific literature. Reagents for research utilizing 3D techniques can be found at www.trevigen.com.

References
1. Birgersdotter, A.; Sandberg, R.; Ernberg, I. Gene expression perturbation in vitro—a growing case for three-dimensional (3-D) culture systems. Semin. Cancer Biol. 2005, 15(5), 405–12.

2. Cukierman, E.; Pankov, R.; Yamada, K.M. Cell interactions with three-dimensional matrices. Curr. Opin. Cell Biol. 2002, 14(5), 633–40.

3. Nelson, C.M.; Bissell, M.J. Of extracellular matrix, scaffolds, and signaling: tissue architecture regulates development, homeostasis, and cancer. Ann. Rev. Cell Dev. Biol. 2006, 22, 287–309.

4. Schuetz, E.G.; Li, D.; Omiecinski, C.J.; Muller-Eberhard, U.; Kleinman, H.K.; Elswick, B.; Guzelian, P.S. Regulation of gene expression in adult rat hepatocytes cultured on a basement membrane matrix. J. Cell Physiol. 1988, 134(3), 309–23.

5. Barcellos-Hoff, M.H.; Aggeler, J.; Ram, T.G.; Bissell, M.J. Functional differentiation and alveolar morphogenesis of primary mammary cultures on reconstituted basement membrane. Development 1989, 105(2), 223–35.

6. Webber, M.M.; Bello, D.; Kleinman, H.K.; Hoffman, M.P. Acinar differentiation by non-malignant immortalized human prostatic epithelial cells and its loss by malignant cells. Carcinogenesis 1997, 18(6), 1225–31.

7. Grant, D.S.; Tashiro, K.; Segui-Real, B.; Yamada, Y.; Martin, G.R.; Kleinman, H.K. Two different laminin domains mediate the differentiation of human endothelial cells into capillary-like structures in vitro. Cell 1989, 58(5), 933–43.

8. Griffith, L.G.; Swartz, M.A. Capturing complex 3-D tissue physiology in vitro. Nat. Rev. Mol. Cell Biol. 2006, 7(3), 211–24.

9. Petersen, O.W.; Ronnov-Jessen, L.; Howlett, A.R.; Bissell, M.J. Interaction with Basement Membrane Serves to Rapidly Distinguish Growth and Differentiation Pattern of Normal and Malignant Human Breast Epithelial Cells. Proceedings of the NationalAcademy of Sciences 1992, 89(19), 9064–8.