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Hum. Reprod. Advance Access originally published online on September 2, 2005
Human Reproduction 2006 21(1):217-222; doi:10.1093/humrep/dei275
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© The Author 2005. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oupjournals.org

Serum-free medium cultivation to improve efficacy in establishment of human embryonic stem cell lines

Peng Hong-mei1,2,3 and Chen Gui-an1,3,4

1 Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing 100083, 2 Department of Obstetrics and Gynecology, General Hospital of PLA, Beijing 100853 and 3 Peking University Stem Cell Research Center, Beijing 100083, China

4 To whom correspondence should be addressed at: Peking University Third Hospital, Department of Obstetrics and Gynecology, Beijing 100083, China. E-mail: chenguian{at}bjmu.edu.cn

BACKGROUND: Serum-containing and serum-free media were used to derive human embryonic stem (HES) cells from donated oocytes and embryos. METHODS and RESULTS: Inner cell masses (ICM) were isolated by immunosurgery. The HES cells were found to be easily obtained and expanded in a serum-free medium. The efficacy in establishing human embryonic stem cell lines improved in a serum-free medium compared with that in serum-containing media. Four HES cell lines were derived from 13 isolated ICM on mouse embryonic fibroblast feeder layers. All four cell lines possess the same characteristics and differentiating potency: normal 46, XX or 46, XY karyotype; and expressing a series of surface markers such as APase, SSEA-3, SSEA-4, TRA-1–60, TRA-1–81, but not SSEA-1. They can form embryoid bodies in suspension culture and develop teratomas comprising derivatives of three embryonic germ layers when injected into severe combined immunodeficient mice. CONCLUSION: These preliminary results suggest that serum-free cultivation may be superior to serum-containing cultivation for deriving human embryonic stem cells.

Key words: embryonic stem cell/derivation/passage/serum-free cultivation


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