National Dairy Research Institute (NDRI), Karnal, India.
PLoS One. 2012;7(7):e40469. doi: 10.1371/journal.pone.0040469. Epub 2012 Jul 9.
The objective of this study was to establish the buffalo mammary epithelial cell line (BuMEC) and characterize its mammary specific functions.
Buffalo mammary tissue collected from the slaughter house was processed enzymatically to obtain a heterogenous population of cells containing both epithelial and fibroblasts cells. Epithelial cells were purified by selective trypsinization and were grown in a plastic substratum. The purified mammary epithelial cells (MECs) after several passages were characterized for mammary specific functions by immunocytochemistry, RT-PCR and western blot.
The established buffalo mammary epithelial cell line (BuMEC) exhibited epithelial cell characteristics by immunostaining positively with cytokeratin 18 and negatively with vimentin. The BuMEC maintained the characteristics of its functional differentiation by expression of β-casein, κ-casein, butyrophilin and lactoferrin. BuMEC had normal growth properties and maintained diploid chromosome number (2n = 50) before and after cryopreservation. A spontaneously immortalized buffalo mammary epithelial cell line was established after 20 passages and was continuously subcultured for more than 60 passages without senescence.
We have established a buffalo mammary epithelial cell line that can be used as a model system for studying mammary gland functions.
本研究旨在建立水牛乳腺上皮细胞系（BuMEC），并鉴定其乳腺特异性功能。
从屠宰场采集水牛乳腺组织，经酶解处理获得含有上皮细胞和成纤维细胞的异质细胞群体。通过选择性胰蛋白酶消化法纯化上皮细胞，并在塑料基质上培养。经过多次传代，通过免疫细胞化学、RT-PCR 和 Western blot 鉴定纯化的乳腺上皮细胞（MECs）的乳腺特异性功能。
建立的水牛乳腺上皮细胞系（BuMEC）通过细胞角蛋白 18 的免疫染色呈阳性，波形蛋白的免疫染色呈阴性，表现出上皮细胞的特征。BuMEC 通过表达β-酪蛋白、κ-酪蛋白、牛乳铁蛋白和乳脂球表皮生长因子 8 保持其功能分化的特征。BuMEC 在冷冻保存前后具有正常的生长特性，并保持二倍体染色体数（2n=50）。经过 20 代传代后建立了一个自发永生化的水牛乳腺上皮细胞系，连续传代超过 60 代而未衰老。
我们已经建立了一个水牛乳腺上皮细胞系，可以作为研究乳腺功能的模型系统。
