草原红牛胎儿成纤维细胞的分离培养及性别鉴定

为了获得高质量的草原红牛胎儿成纤维细胞,试验以发育40 d的草原红牛胎儿为材料,采用组织块贴壁法及胶原酶消化法分离纯化草原红牛胎儿成纤维细胞,并绘制生长曲线,再以SRY基因为模板设计性别鉴定引物,通过对已知性别草原红牛血液基因组PCR扩增检验所设计引物的性别特异性,最后采用基因组PCR方法鉴定分离得到的成纤维细胞的性别。结果表明:成功分离纯化出草原红牛胚胎成纤维细胞,经基因组PCR检验该细胞株性别为雄性。

In vitro culture and sex determination of fetal fibroblast cells from Chinese red steppe

To obtain the high-quality fetal fibroblast(BFFb) cell from Chinese red steppe.The fetal fibroblast cells were isolated and purified from 40 days fetus of Chinese red steppe by tissue explant attachment and collagenase digesting,and the cell growth curve was plotted.The specific primer was designed to amplify SRY gene,which was used to sex determine,the high specificity was proved by genomic PCR using known gender genome as template.In the last,the gender of prepared fetal fibroblast cells were identified by genomic PCR using SRY primer.The results indicated that the fetal fibroblast cells were isolated from Chinese red steppe and the gender of fetal fibroblast cell was identified as the male.