不同方法分离奶牛乳腺上皮细胞体外培养的研究

Study of In Vitro-culture for Bovine Mammary Epithelial Cells with Different Method to Separate

从荷斯坦奶牛乳房无茵采取乳腺组织,通过不同的方法分离、培养、纯化牛乳腺上皮细胞,研究乳腺上皮细胞的体外培养效果。结果表明:组织块接种、0.25%的胰酶和100 u/mL透明质酸酶的混合液37℃消化组织块3 h,可以得到大量细胞用于原代培养。在以DMEM/F12为基础培养液,在培养液中添加10%的胎牛血清、100μg/mL的双抗、表皮生长因子、胰岛素、转铁蛋白、硒酸钠等,组成的完全培养液中奶牛乳腺上皮细胞生长良好。上皮细胞显微结构显示:奶牛乳腺上皮细胞在体外培养过程中含有不同的细胞类型,大多数上皮细胞呈多角形,细胞之间连接成片,细胞界限明显,部分细胞界限不明显。

Researching the effects of the in vitro-culture for bovine mammary epithelial cells （BMEC） with different method to separate, culture and sublime. The results show that the large numbers of BMEC could be separated from mammary tissue inoculation and with 0.25% trypsinase and 100 u/mL hyaluronidase for 3 h at 37 ℃. BMEC grows well in DMEM/F12 culture solution with 10% fetal bovine serum, 100 μg/mL antibiotics, epithelial growth factor （EGF）, insulin, hydrocortisone, transferring and sodium selenate. The light microscope structure indicated that： there are different type cell in vitro, and most of them are guboidal, the cells connected with each other properly, the boundary of the cells is clear, but part of their boundary is not clear.