用囊胚内细胞团建立小鼠胚胎干细胞系的尝试

Establishment of embryonic stem cell line derived from inner cell mass of mouse

目的建立小鼠129品系胚胎干细胞(ES)系。方法分离囊胚内细胞团接种至小鼠胚胎成纤维细胞,培养、传代以获得细胞系,通过形态观察、碱性磷酸酶检查、体内外分化实验予以鉴定。结果共获取12个囊胚,培养后有9个内细胞团增殖,消化传代,分离出2株克隆;形态观察具有明显的ES细胞形态,染色体核型检查为XY,碱性磷酸酶检查细胞呈阳性,体外分化实验表明可自发分化为外、中和内胚层细胞,体内分化实验获得典型的畸胎瘤,并具备小鼠特异性表面标志物Oct-4、SSEA-1。结论应用本方法建立了两株129品系胚胎干细胞系,并证实其符合小鼠胚胎干细胞的特征。

Objective： To establish embryonic stem cell line from mouse strain 129. Methods： Inner cell mass（ICM） was isolated and cultured on the primary mouse embryo fibroblast （PMEF） for the establishment of the stem cell lines. The established embryonic stem cell line was identified by the characteristic morphology, karyotype analysis, alkaline phosphatase（AKP） test, and in vitro and in vivo differentiation capacity. Results： Totally 12 blastocysts were obtained to culture and 9 proliferated ICMs achieved. The ICMs were subject to sequential digestions and subcultures. Two stem cell lines were obtained. The cell lines were confirmed as embryonic stem cell lines according to cell morphology, normal karyotype, alkaline phosphatase test, in vitro and in vivo differentiation capacity test and expression of the surface markers of mouse stem cell. Conclusion： The protocol in our study to establish embryonic stem cell line from mouse strain 129 was feasible.