Culture of human embryonic stem cells

Article metrics

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: Phase contrast images of HESC colonies.
Figure 2: Karyotype and pluripotency testing of the human embryonic stem cell H7.

References

  1. 1

    Heng, B.C., Liu, H. & Cao, T. Feeder cell density--a key parameter in human embryonic stem cell culture. In Vitro Cell Dev. Biol. Anim. 40, 255–257 (2004).

  2. 2

    Lee, J.B. et al. Available human feeder cells for the maintenance of human embryonic stem cells. Reproduction 128, 727–735 (2004).

  3. 3

    Stojkovic, P. et al. An autogeneic feeder cell system that efficiently supports growth of undifferentiated human embryonic stem cells. Stem Cells 23, 306–314 (2005).

  4. 4

    Inzunza, J. et al. Derivation of human embryonic stem cell lines in serum replacement medium using postnatal human fibroblasts as feeder cells. Stem Cells 23, 544–549 (2005).

  5. 5

    Xu, R. et al. Basic FGF and suppression of BMP signalling sustain undifferentiated proliferation of human ES cells. Nat. Methods 2, 185–189 (2005).

  6. 6

    Martin, M.J., Muotri, A., Gage, F. & Varki, A. Human embryonic stem cells express an immunogenic nonhuman sialic acid. Nat. Med. 11, 228–232 (2005).

  7. 7

    Holden, C. Human embryonic stem cells. Getting the mice out of ES cell cultures. Science 307, 1393 (2005).

  8. 8

    Wang, G. et al. Noggin and bFGF cooperate to maintain the pluripotency of human embryonic stem cells in the absence of feeder layers. Biochem. Biophys. Res. Commun. 330, 934–942 (2005).

  9. 9

    Beattie, G.M. et al. Activin A maintains pluripotency of human embryonic stem cells in the absence of feeder layers. Stem Cells 23, 489–495 (2005).

  10. 10

    Xu, C. et al. Basic fibroblast growth factor supports undifferentiated human embryonic stem cell growth without conditioned medium. Stem Cells 23, 315–323 (2005).

  11. 11

    Vallier, L., Reynolds, D. & Pedersen, R.A. Nodal inhibits differentiation of human embryonic stem cells along the neuroectodermal default pathway. Dev Biol 275, 403–421 (2004).

  12. 12

    Thomson, J.A. et al. Embryonic stem cell lines derived from human blastocysts. Science 282, 1145–1147 (1998).

  13. 13

    Pera, M.F., Reubinoff, B. & Trounson, A. Human embryonic stem cells. J. Cell Sci. 113 (Pt. 1), 5–10 (2000).

  14. 14

    Reubinoff, B.E., Pera, M.F., Fong, C.Y., Trounson, A. & Bongso, A. Embryonic stem cell lines from human blastocysts: somatic differentiation in vitro. Nat. Biotechnol. 18, 399–404 (2000).

  15. 15

    Nagy, A., Gersenstein, M., Vintersten, K. & Behringer Manipulating the Mouse Embryo, 371–373 (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2003).

  16. 16

    Pera, M.F., Filipczyk, A.A., Hawes, S.M. & Laslett, A.L. Isolation, characterization, and differentiation of human embryonic stem cells. Methods Enzymol. 365, 429–446 (2003).

  17. 17

    Sambrook, J. & Russell, D.W., Molecular Cloning: A Laboratory Manual, 7.9–7.12 (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA, 2001).

  18. 18

    Simerly, C. & Schatten, G. Techniques for localization of specific molecules in oocytes and embryos. Methods Enzymol. 225, 516–553 (1993).

  19. 19

    Vallier, L. et al. Enhancing and diminishing gene function in human embryonic stem cells. Stem Cells 22, 2–11 (2004).

  20. 20

    Perrier, A.L. et al. Derivation of midbrain dopamine neurons from human embryonic stem cells. Proc. Natl. Acad. Sci. USA 101, 12543–12548 (2004).

  21. 21

    Mummery, C. et al. Cardiomyocyte differentiation of mouse and human embryonic stem cells. J. Anat. 200, 233–242 (2002).

Download references

Acknowledgements

We apologize in advance to the many researchers whose excellent papers could not be referenced, because of space limitations. The methods described in this paper are those presently used at the Pittsburgh Development Center, USA and in the Department of Surgery, Cambridge, UK. They were developed in collaboration with our many good colleagues in the HESC field and in close conjunction with the faculty of our Frontiers in Human Embryonic Stem Cells Course taught annually at the University of Pittsburgh. Also contributing to the preparation and review of this manuscript were many laboratory colleagues, particularly H. Qidwai, J. Mich-Basso and C. Redinger for editorial review.

Author information

Correspondence to Gerald Schatten.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Schatten, G., Smith, J., Navara, C. et al. Culture of human embryonic stem cells. Nat Methods 2, 455–463 (2005) doi:10.1038/nmeth0605-455

Download citation

Further reading