CAJ | 학술논문

目的：制备G418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14 d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G418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G418的筛选培养基中能存活1～2周。
목적：제비G418항성적소서배태간세포자양층。방법：이용일개neo기인성공정합도기인조적소서품계Smad3ex8+/-，종기임신14 d적자서중획취배태，선제비소서배태성섬유세포，PCR감정기인형후，도선일주neo기인잡합형적소서배태성섬유세포，용사렬매소C처리즉획득G418항성적소서배태간세포자양층。결과：이용차방법제비적자양층능유효지지지소서배태간세포적생장，재함G418적사선배양기중능존활1～2주。
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 Smad3ex8+/- 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.