[Establishment of a single cell-cloned mouse embryonic stem cell line].
OBJECTIVE
To establish embryonic stem cells culture system and methods.


METHODS
Inner cell masses were isolated from blastocysts of 3.5 day-old 129SvJ mice by immunosurgery, seeded onto gamma-irradiated mouse fibroblasts feeder layer. The outgrowths were passed by digestion with trypsin. Surface markers were identified by cytochemistry and immunohistochemistry. Karyotype was tested with standard G-banding technique. Differential potency was tested both in vivo and in vitro.


RESULTS
One single cell  cloned mouse embryonic stem cell line G(11) was established, which had normal 40XY karyotype and proliferated rigorously for a long time, expressed surface markers such as AP and SSEA-1, and was able to differentiate into many cell types both in vivo and in vitro.


CONCLUSIONS
Mouse embryonic stem cell culture system has been established in our laboratory.
Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing 100083, China. phmei@tsinghua.edu.cn
Hong-mei
HM
Peng
Gui-an
GA
Chen
D000818:Animals; D001755:Blastocyst / Q000166:cytology*; D018929:Cell Culture Techniques; D002460:Cell Line; D002469:Cell Separation / Q000379:methods; D002999:Clone Cells / Q000166:cytology; D053595:Embryonic Stem Cells / Q000166:cytology*; D005260:Female; D051379:Mice; D008813:Mice, Inbred ICR; D011247:Pregnancy