Establishment and characterization of a lactating bovine mammary epithelial cell model for the study of milk synthesis
Establishment and characterization of a lactating bovine mammary epithelial cell model for the study of milk synthesis
复制标题
用于研究乳汁合成的泌乳牛乳腺上皮细胞模型的建立和表征
DOI：
10.1042/cbi20100023
复制
发表时间：
2010-07-01
影响因子：
3.9
通讯作者：
Liu, Jian-Xin
中科院分区：
文献类型：
--
作者：
Zhao, Ke;Liu, Hong-Yun;Liu, Jian-Xin
本研究旨在建立一种体外泌乳牛乳腺上皮细胞（BMEC）模型，使其能在一段时间内保持天然功能。将泌乳中期荷斯坦奶牛乳腺组织分散培养在含有胰岛素、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。当细胞从组织中迁移到约80%的融合度后，将组织移除，通过0.25%胰蛋白酶反复消化以去除成纤维细胞来富集分泌性上皮细胞。在塑料培养皿中，BMEC细胞呈单层、鹅卵石状、上皮样形态，形成了典型的乳腺上皮细胞的肺泡状结构和岛状单层聚集体。通过抗细胞角蛋白18抗体染色鉴定分离的细胞为上皮细胞。在这些细胞中观察到一个半对数生长曲线和丰富的微绒毛和细胞质脂滴。同时还检测了αs1酪蛋白基因的转录和αs酪蛋白的合成。因此，我们的泌乳BMEC模型可以作为体外研究牛乳腺乳汁合成的有效模型。
This study sought to establish anin 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 ∼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 αs1 casein gene and synthesis of αs 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.