We have previously reported that high concentrations of basic fibroblast growth factor (bFGF) support feeder-independent growth of human embryonic stem (ES) cells, but those conditions included poorly defined serum and matrix components. Here we report feeder-independent human ES cell culture that includes protein components solely derived from recombinant sources or purified from human material. We describe the derivation of two new human ES cell lines in these defined culture conditions.
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Puck, T.T. & Marcus, P.I. Proc. Natl. Acad. Sci. USA 4, 432–437 (1955).
Thomson, J.A. et al. Science 282, 1145–1147 (1998).
Ying, Q.L., Nichols, J., Chambers, I. & Smith, A. Cell 115, 281–292 (2003).
Xu, R.H. et al. Nat. Biotechnol. 20, 1261–1264 (2002).
Xu, R.H. et al. Nat. Methods 2, 185–190 (2005).
Amit, M. et al. Dev. Biol. 227, 271–278 (2000).
Klimanskaya, I. et al. Lancet 365, 1636–1641 (2005).
Xu, C. et al. Stem Cells 23, 315–323 (2005).
Martin, M.J., Muotri, A., Gage, F. & Varki, A. Nat. Med. 11, 228–232 (2005).
Sperger, J.M. et al. Proc. Natl. Acad. Sci. USA 100, 13350–13355 (2003).
Amit, M., Shariki, C., Margulets, V. & Itskovitz-Eldor, J. Biol. Reprod. 70, 837–845 (2004).
Sato, N., Meijer, L., Skaltsounis, L., Greengard, P. & Brivanlou, A.H. Nat. Med. 10, 55–63 (2004).
Watanabe, M., Maemura, K., Kanbara, K., Tamayama, T. & Hayasaki, H. Int. Rev. Cytol. 213, 1–47 (2002).
Takahama, K. et al. Neuropharmacology 25, 339–342 (1986).
Draper, J.S. et al. Nat. Biotechnol. 22, 53–54 (2004).
NIH grants R24-RR017721 and P20-GMO69981 only supported work with federally approved cell lines H1, H7, H9 and H14. New human ES cell derivations and analysis was performed exclusively at WiCell, using privately funded WiCell facilities, equipment and personnel. We thank S. Lindheim for consenting patients who donated their embryos to this research, and D. Faupel for critical reading of the manuscript.
J.A.T. is cofounder and owns stock in Cellular Dynamics International. The remaining authors declare that they have no competing financial interests.
Optimization of physiochemical culture conditions for human ES cells (PDF 1158 kb)
Karyotype analysis of human ES cells cultured in TeSR1 medium (PDF 421 kb)
Pluripotency of human ES cells maintained in TeSR1 medium (PDF 518 kb)
Representative morphology and FACS analysis of human ES cells cultured in defined, feeder-independent conditions. (PDF 604 kb)
Absence of sialic acid contamination in human ES cells cultured in defined conditions (PDF 102 kb)
Karyotype of new human ES cell lines (PDF 212 kb)
Complete formulation for TeSR1 medium (PDF 30 kb)
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Ludwig, T., Levenstein, M., Jones, J. et al. Derivation of human embryonic stem cells in defined conditions. Nat Biotechnol 24, 185–187 (2006). https://doi.org/10.1038/nbt1177
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