【摘要】 本试验采用两种方法制备小鼠胎儿成纤维细胞饲养层(mouse embryon ic fibrob last,MEF),两种方法的差别在于用丝裂霉素-C(m itomyc ine-C,MMC)处理后,一种是用PBS-洗净MMC后直接使用;另一种是丝裂霉素-C处理后,用胰酶消化后重新接种,待细胞融合后再使用。将获得的小鼠胚胎直接接种在MEF上,分别在5~6d后取增殖的ICM团块,消化再接种培养,观察胚胎干细胞(embryon ic stem cell,ES细胞)样集落的生长情况,并对获得的ES细胞样集落进行碱性磷酸酶(AKP)染色鉴定。结果发现两种处理方法得到的MEF培养体系均具有促进ES细胞增殖以及抑制ES细胞的分化的作用,而且得到的ES细胞样集落符合ES细胞的形态特征,AKP呈强阳性。采用本试验的第一种方法的优点是简化了胎儿成纤维细胞饲养层的制备过程,并且适合做ES细胞的饲养层。更多还原

【Abstract】 Two methods were used to prepare the mouse embryonic fibroblast(MEF) feeder layers,and the MEF feeder layers were used to isolate and culture the embryonic stem cells.It was carried out observation to the effect of the various methods on the MEF feeder layers,the effects of the various MEF feeder layers on the development of embryos,the growth of the ICM and the isolation of the embryonic stem(ES)cells,and the optimal passage of the MEF feeder layers were filtered out for preparation of the feeder layers in this experiment.It was find in this experiment that the method for preparation of the MEF feeder layers could simplify the preparation for the MEF feeder layers,but there had no impacts on the isolation the ES cells.更多还原