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Derivation and maintenance of human embryonic stem cells from poor-quality in vitro fertilization embryos

Nature Protocols volume 3, pages 923933 (2008) | Download Citation

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Abstract

Human embryonic stem (hES) cells are self-renewing, pluripotent cells that are valuable research tools and hold promise for use in regenerative medicine. Most hES cell lines are derived from cryopreserved human embryos that were created during in vitro fertilization (IVF) and are in excess of clinical need. Embryos that are discarded during the IVF procedure because of poor morphology and a low likelihood for generating viable pregnancies or surviving the cryopreservation process are also a viable source of hES cells. In this protocol, we describe how to derive novel hES cells from discarded poor-quality embryos and how to maintain the hES cell lines.

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Acknowledgements

This work was made possible through the generosity and vision of Joshua and Anita Bekenstein, the Harvard Stem Cell Institute and Children's Hospital Boston. G.Q.D. is a recipient of the Burroughs Wellcome Fund Clinical Scientist Award in Translational Research.

Author information

Affiliations

  1. Division of Newborn Medicine, Brigham & Women's Hospital and Children's Hospital Boston, Karp Family Research Building 7214, 300 Longwood Avenue, Boston, Massachusetts 02115, USA.

    • Paul H Lerou
  2. Division of Pediatric Hematology Oncology, Children's Hospital Boston, 300 Longwood Avenue, Boston, Massachusetts 02115, USA.

    • Akiko Yabuuchi
    • , Hongguang Huo
    • , Justine D Miller
    • , Leah F Boyer
    • , Thorsten M Schlaeger
    •  & George Q Daley
  3. Harvard Stem Cell Institute, hES Cell Core Facility, Children's Hospital Boston, Karp Family Research Building 09213, 300 Longwood Avenue, Boston, Massachusetts 02115, USA.

    • Hongguang Huo
    • , Justine D Miller
    • , Leah F Boyer
    • , Thorsten M Schlaeger
    •  & George Q Daley
  4. Division of Hematology, Brigham and Women's Hospital, Karp Family Research Building 7214, 300 Longwood Avenue, Boston, Massachusetts 02115, USA.

    • George Q Daley
  5. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Karp Family Research Building 7214, 300 Longwood Avenue Boston, Massachusetts 02115, USA.

    • George Q Daley
  6. Harvard Stem Cell Institute, Karp Family Research Building 7214, 300 Longwood Avenue, Boston, Massachusetts 02115, USA.

    • George Q Daley
  7. Howard Hughes Medical Institute, Boston, Massachusetts 02115, USA.

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Contributions

P.H.L. and A.Y. contributed equally to this work.

Corresponding author

Correspondence to George Q Daley.

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DOI

https://doi.org/10.1038/nprot.2008.60

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