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Derivation of human embryonic stem cells from single blastomeres

Nature Protocols volume 2, pages 19631972 (2007) | Download Citation

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Abstract

This protocol details a method to derive human embryonic stem (hES) cells from single blastomeres. Blastomeres are removed from morula (eight-cell)-stage embryos and cultured until they form multicell aggregates. These blastomere-derived cell aggregates are plated into microdrops seeded with mitotically inactivated feeder cells, and then connected with neighboring microdrops seeded with green fluorescent protein-positive hES cells. The resulting blastomere-derived outgrowths are cultured in the same manner as blastocyst-derived hES cells. The whole process takes about 3–4 months.

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Acknowledgements

We thank Tong Li and Marc Maserati for help with cell culture, karyotyping and embryo manipulations.

Author information

Affiliations

  1. Advanced Cell Technology, 381 Plantation Street, Worcester, Massachusetts 01605, USA.

    • Irina Klimanskaya
    • , Young Chung
    • , Sandy Becker
    • , Shi-Jiang Lu
    •  & Robert Lanza

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Competing interests

The authors are employees of Advanced Cell Technology, a biotechnology company in the field of stem cell research and regenerative medicine.

Corresponding author

Correspondence to Robert Lanza.

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DOI

https://doi.org/10.1038/nprot.2007.274

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