Abstract
Embryonic stem (ES) cells have the ability to self-renew as well as differentiate into any cell type in the body. These traits make ES cells an attractive “raw material” for a variety of cell-based technologies. However, uncontrolled cell aggregation in ES cell differentiation culture inhibits cell proliferation and differentiation and thwarts the use of stirred suspension bioreactors. Encapsulation of ES cells in agarose microdrops prevents physical interaction between developing embryoid bodies (EBs) that, in turn, prevents EB agglomeration. This enables use of stirred suspension bioreactors that can generate large numbers of ES-derived cells under controlled conditions.
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References
Keller, G. M. (1995) In vitro differentiation of embryonic stem cells. Curr. Opin. Cell Biol. 7, 862–869.
Boheler, K. R., Czyz, J., Tweedie, D., Yang, H. T., Anisimov, S. V., and Wobus, A. M. (2002) Differentiation of pluripotent embryonic stem cells into cardiomyocytes. Circ. Res. 91, 189–201.
Soria, B., Roche, E., Berna, G., Leon-Quinto, T., Reig, J. A., and Martin, F. (2000) Insulin-secreting cells derived from embryonic stem cells normalize glycemia in streptozotocin-induced diabetic mice. Diabetes 49, 157–162.
Lee, S. H., Lumelsky, N., Studer, L., Auerbach, J. M., and McKay, R. D. (2000) Efficient generation of midbrain and hindbrain neurons from mouse embryonic stem cells. Nature Biotechnol. 18, 675–679.
Kaufman, D. S., Hanson, E. T., Lewis, R. L., Auerbach, R., and Thomson, J. A. (2001) Hematopoietic colony-forming cells derived from human embryonic stem cells. Proc. Natl. Acad. Sci. USA 98, 10,716–10,721.
Keller, G., Kennedy, M., Papayannopoulou, T., and Wiles, M. V. (1993) Hematopoietic commitment during embryonic stem cell differentiation in culture. Mol. Cell. Biol. 13, 473–486.
Dang, S. M., Kyba, M., Perlingeiro, R., Daley, G. Q., and Zandstra, P. W. (2002) Efficiency of embryoid body formation and hematopoietic development from embryonic stem cells in different culture systems. Biotechnol. Bioeng. 78, 442–453.
Lame, L., Antos, C., Butz, S., et al. (1996) A role for cadherins in tissue formation. Development 122, 3185–3194.
Viswanathan, S., Benatar, T., Rose-John, S., Lauffenburger, D. A., and Zandstra, P. W. (2002) Ligand/receptor signaling threshold (LIST) model accounts for gp130-mediated embryonic stem cell self-renewal responses to LIF and HIL-6. Stem Cells 20, 119–138.
Zandstra, P. W. and Nagy, A. (2001) Stem cell bioengineering. Annu. Rev. Biomed. Eng. 3, 275–305.
Dang, S. M., Gerecht-Nir, S., Chen, J., Itskovitz-Eldor, J., and Zandstra, P. W. (2004) Controlled, scalable embryonic cell differentiation culture. Stem Cells 22, 275–282.
Weaver, J. C., McGrath, P., and Adams, S. (1997) Gel microdrop technology for rapid isolation of rare and high producer cells. Nature Med. 3, 583–585.
Adelman, C. A., Chattopadhyay, S., and Bieker, J. J. (2002) The BMP/BMPR/Smad pathway directs expression of the erythroid-specific EKLF and GATA1 transcription factors during embryoid body differentiation in serum-free media. Development 129, 539–549.
Chadwick, K., Wang, L., Li, L., et al. (2003) Cytokines and BMP-4 promote hematopoietic differentiation of human embryonic stem cells. Blood 102, 906–915.
Turksen, K. (ed.) (2002) Embryonic Stem Cells: Methods and Protocols, Humana, Totowa, NJ.
Chiu, A. and Rao, M. S. (eds.) (2003) Human Embryonic Stem Cells, Humana, Totowa, NJ.
Magyar, J. P., Nemir, M., Ehler, E., Suter, N., Perriard, J. C., and Eppenberger, H. M. (2001) Mass production of embryoid bodies in microbeads. Ann. NY Acad. Sci. 944, 135–143.
Gin, H., Dupuy, B., Baquey, C., Ducassou, D., and Aubertin, J. (1987) Agarose encapsulation of islets of Langerhans: reduced toxicity in vitro. J. Microencapsul. 4, 239–242.
Tashiro, H., Iwata, H., Tanigawa, M., et al. (1998) Microencapsulation improves viability of islets from CSK miniature swine. Transplant. Proc. 30, 491.
Stevenson, W. T., Evangelista, R. A., Sugamori, M. E., and Sefton, M. V. (1988) Microencapsulation of mammalian cells in a hydroxyethyl methacrylate-methyl methacrylate copolymer: preliminary development. Biomater. Artif. Cells Artif. Organs 16, 747–769.
Dupuy, B., Gin, H., Baquey, C., and Ducassou, D. (1988) In situ polymerization of a microencapsulating medium round living cells. J. Biomed. Mater. Res. 22, 1061–1070.
Zekorn, T., Siebers, U., Horcher, A., et al. (1992) Alginate coating of islets of Langerhans: in vitro studies on a new method for microencapsulation for immunoisolated transplantation. Acta Diabetol. 29, 41–45.
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Dang, S.M., Zandstra, P.W. (2005). Scalable Production of Embryonic Stem Cell-Derived Cells. In: Helgason, C.D., Miller, C.L. (eds) Basic Cell Culture Protocols. Methods in Molecular Biology™, vol 290. Humana Press. https://doi.org/10.1385/1-59259-838-2:353
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DOI: https://doi.org/10.1385/1-59259-838-2:353
Publisher Name: Humana Press
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