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Derivation of human embryonic stem cells in defined conditions


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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Figure 1: Evaluation of TeSR1-cultured human ES5 cells.
Figure 2: Representative morphology, RT-PCR and FACS analysis of human ES cells derived and maintained in defined, feeder-independent culture conditions.

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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.

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Correspondence to James A Thomson.

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J.A.T. is cofounder and owns stock in Cellular Dynamics International. The remaining authors declare that they have no competing financial interests.

Supplementary information

Supplementary Fig. 1

Optimization of physiochemical culture conditions for human ES cells (PDF 1158 kb)

Supplementary Fig. 2

Karyotype analysis of human ES cells cultured in TeSR1 medium (PDF 421 kb)

Supplementary Fig. 3

Pluripotency of human ES cells maintained in TeSR1 medium (PDF 518 kb)

Supplementary Fig. 4

Representative morphology and FACS analysis of human ES cells cultured in defined, feeder-independent conditions. (PDF 604 kb)

Supplementary Fig. 5

Absence of sialic acid contamination in human ES cells cultured in defined conditions (PDF 102 kb)

Supplementary Fig. 6

Karyotype of new human ES cell lines (PDF 212 kb)

Supplementary Table 1

Complete formulation for TeSR1 medium (PDF 30 kb)

Supplementary Methods (DOC 72 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).

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