用饲养层分离胚胎干细胞集落的实验初探

目的 用饲养层分离胚胎干细胞集落。方法 用胚龄为13~14 d的小鼠胚胎分离原代成纤维细胞，制成饲养层，用于囊胚的培养。结果 小鼠原代胚胎成纤维细胞（PMEF）贴壁能力较好，增殖快，易铺层。囊胚和内细胞团（ICM）在饲养层上贴壁生长良好，当培养4~5 d时，其增殖率为16/28（57%）。在ICM离散48 h后，各种胚胎干细胞（ES）集落开始出现。此种集落经碱性磷酸酶染色成阳性。结论 用饲养层分离胚胎干细胞获得初步成功。

Isolation of ES Cell Colonies on a Feeder Layer of ICR Mouse Fibroblasts

Objective To isolate ES cell colonies on a feeder layer. Methods Mouse embryos of 13-14 days were for the cultivation of primary mouse embryo fibroblasts (PMEF) which turned to be a feeder layer for blastocysts. Results The inner cell mass (ICM) of blastocysts grew well on the feeder layer of PMEF.The proliferation rate of ICM was 57%. After 48 hours of ICM disagregation ES colonies began to appear,showing strong AKP activity. Conclusion It is a preliminary success for the feeder layer to produce embryonic stem cell lines.