不同性别胎鼠生殖嵴表观特征差异性研究

Variations of Epigenetic Characteristics of Germinal Ridge between Male Embryos and Female Embryos

在应用小鼠胚胎干细胞(ESCs)裂解液诱导异性小鼠胚胎成纤维细胞(MEFs)为诱导性多能干细胞(iPSCs)的过程中,通过性别鉴定的方法筛选特定性别的被诱导的MEFs,能够快速简易的鉴定出iPSCs的生成材料是否来源于被诱导的体细胞。利用妊娠13.5天雄性和雌性胎鼠生殖嵴在形态学上存在明显差异的特性,识别出不同性别昆明系小鼠13.5天的胚胎,并从中分离培养出大量不同性别的MEFs,用于iPSCs的实验研究。根据雄性小鼠特异性的Y染色体性别决定区(SRY基因)的核心序列,设计一对引物,并对获得的不同性别MEFs的DNA样本进行PCR扩增,验证不同性别胎鼠生殖嵴在形态学上存在的差异。结果表明,PCR性别鉴定结果与MEFs来源胚胎的性别一致。不同性别的MEFs均具有典型的成纤维细胞特性,两者在形态学上并没有差异。因此,利用雄性与雌性胚胎在生殖嵴形态学上的差异可以快速分离获得大量不同性别的MEFs。

When mouse embryonic fibroblasts （MEFs） co-incubated with extracts of embryonic stem cells （ESCs） to reprogram into the induced pluripotent stem cells （iPSCs）, the origin of iPSCs could be rapidly and easily identified based on the different gender of MEFs and embryonic stem cells. Different sexual MEFs were isolated from different sexual Kunming mouse embryos at 13.5 dpc （days post coitum） which were identified according to the morphological differences between the male germinal ridge and the female germinal ridge. A pair of primers were designed according to the core gene sequence of special sex-determining region of Y-chromosome （SRY gene） of male mouse, and DNA samples obtained from different sexual MEFs were amplified by PCR to test and verify the differences in morphology between the different sexual germinal ridge. The results indicated that the genderof MEFs which were identified with sex identification with PCR was consistent with that of mouse embryos at 13.5 dpc. Different sexual MEFs manifested the typical characteristics of fibroblasts, and there were no difference in morphology between the male MEFs and the female MEFs. Therefore, a large number of different sexual MEFs could be isolated from different sexual mouse embryos at 13.5 dpc which were identified according to the morphological difference between the male germinal ridge and the female germinal ridge.