牦牛乳腺上皮细胞系的建立与生物学特性  

Establishment and characterization of yak mammary epithelial cell line

作　　者：郝明超[1] 刘犇[1] 樊江峰[1] 陈鹏[1] 张华[1] 王明亮[1] 高旭东[1] 余四九[1] 

机构地区：[1]甘肃农业大学动物医学院,兰州730070

出　　处：《兽类学报》2013年第2期157-163,共7页Acta Theriologica Sinica

基　　金：国家科技支撑计划(2009BAC53B06)

摘　　要：本研究旨在建立牦牛乳腺上皮细胞体外培养体系。采用胶原酶消化法成功地建立了牦牛乳腺上皮细胞系(YMEC),通过免疫细胞化学、超微结构观察和RT-PCR法对YMEC细胞进行了鉴定,并研究了其形态、活力、生长曲线以及核型等生物学特性。结果表明,YMEC细胞染色体2n=60,群体倍增时间为45～48h,持续培养25代后出现细胞分化;细胞呈典型的"铺路石样"形态,其表面有丰富的微绒毛,细胞质内含丰富的线粒体和粗面内质网。污染检测结果为阴性。在激素诱导培养时,检测到了β-酪蛋白mRNA的表达。表明本研究成功建立了保留泌乳功能的牦牛乳腺上皮细胞系,为研究牦牛乳腺上皮细胞的功能提供了理想的工具。The present study was carried out to establish a method of cultivation of yak mammary epithelial cell in vitro. Yak mammary epithelial cell line （YMEC） was established successfully by use of collagenase digestion, and then identi- fied by immunocytochemistry, ultrastructural characteristics and RT-PCR. The morphological character, doubling time of cell population, growth curve, and chromosomal analysis of YMEC were characterized. The results showed that YMEC maintained a normal diploid chromosome modal number of 2n = 60, possessed an estimated population doubling time of 45 - 48 h and differentiation appeared after subcuhuring 25 times. The cell showed typical ＇ cobblestone＇ morphology with numerous microvilli on its surface and there existed abundant amounts of rough endoplasmic reticulum and mitochondria in its cytoplasm. The result of sterility was negative. When the cells were stimulated by hormones, mRNA of beta-casein was detected. It was concluded that YMEC line retaining mammary gland specific functions was established, which might be a useful tool for studying the function of yak mammary epithelial cell.

关 键 词：牦牛 乳腺上皮细胞 细胞系 生物学特性 

分 类 号：Q492[生物学—生理学]