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Derivation of human embryonic stem cells in defined conditions

Nature Biotechnology volume 24, pages 185187 (2006) | Download Citation

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Abstract

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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Acknowledgements

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.

Author information

Affiliations

  1. WiCell Research Institute, PO Box 7365, Madison, Wisconsin 53707-7365, USA.

    • Tenneille E Ludwig
    • , Mark E Levenstein
    • , Jeffrey M Jones
    • , W Travis Berggren
    • , Erika R Mitchen
    • , Leann J Crandall
    • , Kevin R Conard
    • , Marian S Piekarczyk
    • , Rachel A Llanas
    •  & James A Thomson
  2. The Genome Center of Wisconsin, 425 Henry Mall, Madison, Wisconsin 53706, USA.

    • Tenneille E Ludwig
    • , Jennifer L Frane
    • , Christine A Daigh
    •  & James A Thomson
  3. The Wisconsin National Primate Research Center, University of Wisconsin-Madison, 1220 Capitol Court, Madison, Wisconsin 53715-1299, USA.

    • Tenneille E Ludwig
    • , Jennifer L Frane
    • , Christine A Daigh
    •  & James A Thomson
  4. Department of Obstetrics and Gynecology, University of Wisconsin-Madison Medical School, 600 Highland Avenue, Madison, Wisconsin 53792-6188, USA.

    • Jeffrey M Jones
    •  & James A Thomson
  5. The Department of Anatomy, University of Wisconsin-Madison Medical School, 470 N. Charter Street, Madison, Wisconsin 53706-1509, USA.

    • James A Thomson

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Competing interests

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

Corresponding author

Correspondence to James A Thomson.

Supplementary information

PDF files

  1. 1.

    Supplementary Fig. 1

    Optimization of physiochemical culture conditions for human ES cells

  2. 2.

    Supplementary Fig. 2

    Karyotype analysis of human ES cells cultured in TeSR1 medium

  3. 3.

    Supplementary Fig. 3

    Pluripotency of human ES cells maintained in TeSR1 medium

  4. 4.

    Supplementary Fig. 4

    Representative morphology and FACS analysis of human ES cells cultured in defined, feeder-independent conditions.

  5. 5.

    Supplementary Fig. 5

    Absence of sialic acid contamination in human ES cells cultured in defined conditions

  6. 6.

    Supplementary Fig. 6

    Karyotype of new human ES cell lines

  7. 7.

    Supplementary Table 1

    Complete formulation for TeSR1 medium

Word documents

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    Supplementary Methods

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Publication history

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DOI

https://doi.org/10.1038/nbt1177

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