Serum-free medium cultivation to improve efficacy in establishment of human embryonic stem cell lines.
BACKGROUND
Serum-containing and serum-free media were used to derive human embryonic stem (HES) cells from donated oocytes and embryos.


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.


CONCLUSIONS
These preliminary results suggest that serum-free cultivation may be superior to serum-containing cultivation for deriving human embryonic stem cells.
Hong-mei
Peng
P
Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing 100083, China.
Gui-an
Chen
C
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line / Q000166:cytology / Q000502:physiology*; D016895:Culture Media, Serum-Free; D004622:Embryo, Mammalian / Q000166:cytology; D006801:Humans; D007621:Karyotyping; D051379:Mice; D016513:Mice, SCID; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology / Q000502:physiology*
D016895:Culture Media, Serum-Free