牦牛输卵管上皮细胞分离培养和纯化鉴定

Study on isolation,culture and identification of yak oviduct epithelium cells

为建立牦牛输卵管上皮细胞原代培养及纯化方法,通过选取牦牛输卵管,运用机械刮取法和0.25%胰蛋白酶消化两种方法分离上皮细胞进行体外培养。对不同分离方法的培养效果比较,培养细胞进行形态学观察与传代培养、MTT比色检测细胞活力并制定生长曲线,原代及传代上皮细胞的免疫组织化学鉴定,冷冻解冻后经台盼蓝排斥试验检测活细胞数。结果表明该试验分离出的原代细胞,纯化后传代培养,经鉴定为牦牛输卵管上皮细胞,培养的细胞生长状况良好,建立了一套牦牛输卵管上皮细胞分离培养及纯化鉴定的方法。

To study the suitable method for isolation and culture of yak oviduct epithelial cells,the primary epithelial cells were obtained from yak oviduct by mechanical scraping method and 0. 25% trypsin digestion. Culutred primary epithelial cells and passage cells were identified by their culture method,morphological changes,growth curve,immunocytochemistry and cell viability after thawing. Results showed that the yak primary oviduct epithelial cells were isolated successfully. Subculture of primary epithelial cells can be performed after purification and cell growth in good condition. At the same time,the culture model for isolation,culture and identification of yak oviduct epithelium cells was established in vitro.