麦洼牦牛乳腺上皮细胞系的建立及其生物学特性

The Establishment of Mammary Gland Epithelial Cell Line and Its Biological Characteristic Research in Maiwa Yak

为建立麦洼牦牛乳腺上皮细胞的体外培养方法,采用组织块贴壁法及Ⅰ型胶原酶消化法,从麦洼牦牛乳腺组织中成功分离并获得纯化的乳腺上皮细胞,并利用细胞角蛋白18的免疫荧光染色对乳腺上皮细胞进行鉴定。通过细胞生长曲线、群体倍增时间、细胞接种存活率、细胞活力等生物学特性的检测,建立并鉴定麦洼牦牛乳腺上皮细胞系。结果显示,乳腺细胞形态良好,细胞接种存活率达91%,群体倍增时间26.97h,细胞生长曲线呈典型"S"型,细胞传至25代以上仍保持旺盛的增殖活力。可见,通过对细胞传代及反复冻存和复苏已成功建立麦洼牦牛乳腺上皮细胞系,获得大量的乳腺上皮细胞,使麦洼牦牛物种在细胞水平上得以保存,为建立牦牛乳腺生物反应器及泌乳调控机制研究提供基础。

In order to develop a method of growing the Maiwa Yak mammary gland epithelial cells in vitro, tissue mass inoculation and collagenase digestion method were used in this study. Mammary ep- ithelial cells of Maiwa Yak were successfully cultivated and mammary gland epithelial cells were iden- tified by cytokeratin 18 immunofluorescence staining. The survival rate of cell inoculation, doubling time of cell population, growth curve, and morphological and biological characters were measured, and identification and establishment of Maiwa Yak mammary epithelial cell lines were implemented. The results showed that survival efficiency of cell planting was 91 ~ and population doubling time of cells was 26.97 hours. Cells exhibited robust proliferation capability after 25 generations. Through the cell passage and repeatedly freeze-thaw processes, Maiwa yak mammary gland epithelial cell lines were established, and a large number of mammary epithelial cells was acquired. Therefore, Maiwa Yak, an important genetic resource, was preserved in cell level. These results provided basis for the study of lactation mechanism and mammary gland bioreactor of Yak.