南京医科大学学报（自然科学版）2011，Vol.31Issue(3)：295-300,6.
无bFGF条件下简便有效建立C57BL/6J小鼠胚胎干细胞系
Efficient method to establish C57BL/6J mouse embryonic stem cell lines with bFGF-free knockout serum replacement
摘要
Abstract
Objective:To establish C57BL/6J mouse embryonic stem(ES) cell lines with bFGF-free knockout serum replacement (KSR). Metbods:C57BL/6J mouse blastocysts 3.5 days post coition (dpe) were collected and cultured in the medium supplemented with KSR and 1 000 U/mL LIF, with or without 10 ng/mL bFGF. The ES cell lines were identificated by morphology, alkaline phosphatase activity, karyotype, cell surface maker SSEA-1 and pluripotent markers(Oct4, Sox2 and Nanog) analysis. The differentiation ability of ES cells was analyzed both in vitro (embryoid bodies) and in vivo (teratoma). Results: Four cell lines were established out of 6 blastocysts cultured in the medium without bFGF; in contrast, no ES-like colony was observed when bFGF was added to the medium after isolation of 3 high quality blastocysts. The ES cells proliferated well without evidence of differentiation.The ES cells had typical ES-like morphology, expressed alkaline phosphatase,SSEA-1, pluripotent makers such as Oct4, Sox2 and Nanog. They could form embryoid bodies when been cultured in suspension and generate teratomas when been injected subcutaneously into SCID mice, consistent with the the pluripotent abilities of ES cells. Conclusion:KSR containing medium without bFGF could be an efficient culture system for the establishment of mouse ES cell lines.关键词
胚胎干细胞/碱性成纤维细胞生长因子/knockout血清替代品/C57BL/6J小鼠Key words
embryonic stem cells/ basic fibroblast growth factor/ knockout serum replacement/ C57BL/6J mouse分类
生物科学引用本文复制引用
王晏鹏,蒋春艳,董娟,陈娟,高莉,廖婷婷,崔毓桂,钱晓乔,刘嘉茵..无bFGF条件下简便有效建立C57BL/6J小鼠胚胎干细胞系[J].南京医科大学学报（自然科学版）,2011,31(3):295-300,6.基金项目
江苏省医学重点学科资助(XK200702) （XK200702）
