昆明白小鼠胚胎干细胞分离与体外培养

为探索昆明白小鼠胚胎干细胞建系方法,将受孕4.5天的昆明白小鼠囊胚用免疫手术法去除滋胚层,然后将内细胞团(ICM)接种于胎鼠成纤维细胞饲养层上培养,形成的胚胎干细胞样集落用胰蛋白酶-EDTA消化法传代,培养后进行相差显微镜观察及碱性磷酸酶染色。结果饲养层上生长的ICM细胞呈典型的ES样细胞集落,传至第8代碱性磷酸酶染色呈强阳性。实验表明免疫手术法适用于昆明白小鼠ES细胞建系,获得的细胞集落具有ES细胞的主要生物学性状。

Isolation and Culture of Mouse Embryonic Stem Cells of Kunming Species in Vitro

To investigate the method of establishing embryonic stem (ES) cell line from Kunming species mouse, inner cell masses (ICMs) isolated by immunosurgery from 4.5 d blastocysts of Kunming species mouse were cultured on the fetal mouse fibroblast cell feeder layers. The ES cell-like colones were passaged by trypsin- EDTA digest method to 8th generation. Identified by phasecontrast microscope observation and AKP staining, the obtained ES-like cells proved to be AKP intensively positive and able to form typical ES-like colones. This research confirmed that immunosurgery is suitable for establishing ES cell lines from Kunming species mouse and the cells obtained still maintain the main characteristics of ES cell.