摘要
目的 :制备G4 18抗性的小鼠胚胎干细胞滋养层。方法 :利用一个neo基因成功整合到基因组的小鼠品系Smad3ex8+/ ,从其妊娠 14d的雌鼠中获取胚胎 ,先制备小鼠胚胎成纤维细胞 ,PCR鉴定基因型后 ,挑选一株neo基因杂合型的小鼠胚胎成纤维细胞 ,用丝裂霉素C处理即获得G4 18抗性的小鼠胚胎干细胞滋养层。结果 :利用此方法制备的滋养层能有效地支持小鼠胚胎干细胞的生长 ,在含G4 18的筛选培养基中能存活 1～
Objective:To prepare a kind of G418 resistance fibroblast feeder layers.Methods: In order to prepare a stock primary embryonic fibroblast (EMFI) cells, the pregnant Smad3 ex8+/ mice containing heterozygous neo gene were killed 14 days after coitus to get the embryo. EMFI cells were tested by PCR to identify their gene types. Only one EMFI cell which contained heterozygous neo gene was chosen and treated with mitomycin C.The mitomycin C treated EMFI feeders were stored at -70℃. Results: The EMFI feeder layers could support the growth of embryonic stem cell line TC 1 efficiently and could live in G418 resistance medium for 1-2 weeks. [
出处
《军事医学科学院院刊》
CSCD
北大核心
2001年第1期59-61,共3页
Bulletin of the Academy of Military Medical Sciences
基金
国家高技术研究发展计划! (10 2 0 8 0 8 0 2 )
自然科学基金! (39970 413)资助
