Institute of Dairy Science, MOE Key Laboratory of Molecular Animal Nutrition, Zhejiang University, No. 268 Kaixuan Road, Hangzhou 310029, Peoples Republic of China.
Cell Biol Int. 2010 Jul;34(7):717-21. doi: 10.1042/CBI20100023.
This study sought to establish an in vitro lactating BMEC (bovine mammary epithelial cell) model, which may maintain the native function for a period of time. Mammary tissues of midlactation Holstein dairy cows were dispersed and cultured in a medium containing insulin, prolactin, hydrocortisone, transferrin, epidermal growth factor and fetal calf serum. After the cells migrating from the tissue reached approximately 80% of confluency, the tissues were removed, and secretory epithelial cells were enriched by digesting with 0.25% trypsin repeatedly to remove fibroblasts. The BMEC cells plated on plastic dishes displayed a monolayer, cobblestone, epithelial-like morphology and formed alveoli-like structures and island monolayer aggregates which are the typical characteristics of the mammary epithelial cells. The isolated cells were identified as of epithelial origin by staining with antibody against cytokeratin 18. A one-half logarithmically growth curve and abundant microvilli and cytoplasmic lipid droplets were observed in these cells. The transcription of the alphas1 casein gene and synthesis of alphas caseins were also detected in the model. Thus, our lactating BMEC model can be an effective model in vitro for studies of milk synthesis in the bovine mammary gland.
本研究旨在建立一种体外泌乳期牛乳腺上皮细胞 (BMEC) 模型，该模型可以在一段时间内保持天然功能。从中期泌乳荷斯坦奶牛的乳腺组织中分离出来，在含有胰岛素、催乳素、氢化可的松、转铁蛋白、表皮生长因子和胎牛血清的培养基中进行培养。当组织中的细胞迁移到约 80%融合时，将组织去除，并通过反复用 0.25%胰蛋白酶消化来去除成纤维细胞，从而富集分泌上皮细胞。在塑料培养皿上培养的 BMEC 细胞呈单层、鹅卵石状、上皮样形态，并形成类似于肺泡的结构和岛状单层聚集，这些都是乳腺上皮细胞的典型特征。通过对细胞角蛋白 18 抗体的染色，鉴定这些分离的细胞为上皮来源。这些细胞中观察到了半对数生长曲线和丰富的微绒毛和细胞质脂滴。在该模型中还检测到了αs1 酪蛋白基因的转录和αs 酪蛋白的合成。因此，我们的泌乳期 BMEC 模型可以成为研究牛乳腺中乳汁合成的有效体外模型。
Zotero 插件