组织块种植法体外培养奶牛乳腺上皮细胞

Explants culture of cow mammary epithelial cells in vitro

为建立成熟的奶牛乳腺上皮细胞体外培养体系,以组织块种植法培养荷斯坦泌乳期成年母牛乳腺组织,观测长出的上皮细胞在各生长时期的形态变化,绘制生长曲线并计算群体倍增时间,用免疫组化技术鉴定乳腺上皮细胞角蛋白18的表达,用SDS-PAGE电泳和免疫印迹分析技术检测乳腺细胞的酪蛋白分泌情况。试验结果表明,此培养体系可获得良好的奶牛乳腺上皮细胞,原代以及传代培养,细胞增殖旺盛,长势良好;细胞骨架蛋白——角蛋白18表达为阳性,且培养的细胞具有分泌牛乳酪蛋白的功能。

To establish an in vitro culture system in which cow mammary epithelial cells（MEC）grow well,the mammary tissue clumps from Holstein cows in lactation period at adulthood were obtained and cultured to harvest epithelial cells.The cell morphological varieties,cell population doubling time,and the cell growth curve were observed and investigated.The expression of MEC protein 18 of MEC was authenticated using immunity histochemistry technique.The lactation function of the cells was detected by SDS-PAGE and Western-blot.The results showed that the cow MEC grew well and proliferated vigorously in this culture system.The celluar skeletal protein 18 expressed positively,and the electrophoresis and Western-blot analysis of the products of the cells showed that the cells cultured had a function of secreting cow caseins.