山羊乳腺上皮细胞培养体系的建立

Establishment of culture system in goat mammary epithelial cell

为研究乳腺上皮细胞增殖和分化特性及其基因表达的分子机制 ,建立了山羊乳腺上皮细胞培养体系。用无菌手术法从健康第二胎泌乳期山羊乳腺取得组织块 ,在体外条件下用组织块法进行原代培养。基础培养液用 RPMI1 6 4 0 ,含胎牛血清 1 5 0 m L/ L,添加 EGF(1 0 μg/ L)、胰岛素 (0 .1 m g/ L)、E2 (1 μg/ L)、氢化可的松 (0 .1mg/ L)、孕酮 (1 m g/ L)、青霉素 (0 .1 g/ L)及链霉素 (0 .0 5 g/ L) ,在铺有胶原的塑料培养板上培养。从细胞接种存活率、细胞群体倍增时间、细胞生长曲线和细胞分裂指数 4个方面考察了乳腺上皮细胞的生物学性状。结果表明 ,在此培养体系中 ,山羊乳腺上皮细胞生长良好 ,增殖旺盛。细胞产物电泳及蛋白印迹分析结果表明 ,培养的细胞为山羊乳腺上皮细胞 。

Culture system in goat mammary epithelial cell was established to study the proliferation and differentiation and molecular mechanism of gene express in mammary epithelial cell.The mammary tissues from healthy goat second born duration of lactation have been cultured in vitro .The basic medium is RPIM1640 and 15% fetal calf serum,and EGF (10 μg/L),insulin (0.1 mg/L),E 2 (1 μg/L),hydrocortisone (0.1 mg/L),P (1 mg/L),penicillin (0.1 g/L) and streptomycin (0.05 g/L) were added.The cell cultured on the plastics that covered collagen.The cell biological properties were observed and investigated from the cell planting survival efficiency,cell population doubling time,cell growth curve and cell division index.The results showed that the goat mammary epithelial cell grows well and proliferates vigorously in this culture system.The electrophoresis and western blot of the products of the cell showed that the cultured cell is goat mammary epithelial cell,and it has a function that expresses the goat casein.