奶牛乳腺上皮细胞体外培养体系的建立  

Establishment of Bovine Mammary Epithelial Cell Culture System

作　　者：崔立莉[1] 卜登攀[1] 王加启[1] 罗淑萍[2] 

机构地区：[1]中国农科院畜牧所动物营养学国家重点实验室,北京100094 [2]新疆农业大学农学院,新疆乌鲁木齐830052

出　　处：《新疆农业科学》2006年第5期394-398,F0002,共6页Xinjiang Agricultural Sciences

基　　金：国家科技部"十五"科技攻关计划"奶业重大关键技术研究与产业化技术集成示范"(2002BA51802)

摘　　要：通过乳汁中提取细胞进行传代培养获得增殖且可分泌特异性酪蛋白的奶牛乳腺上皮细胞,细胞形态以鹅卵石型和多角型为主,逐渐趋于多角型。当多角型细胞增殖融合到一定阶段,会出现乳头状和圆顶型结构。并进一步对泌乳中后期乳汁分离的细胞做细胞生物学性状检测:细胞接种存活率达到81%;细胞生长曲线呈典型的“S”形,符合细胞生长的一般生物学规律;细胞群体倍增时间最短达48.7 h;经过对乳腺上皮细胞特异性分泌蛋白-酪蛋白的检测,证明培养获得的细胞为乳腺上皮细胞,且具有正常分泌乳蛋白功能。试验初步建立了奶牛乳腺上皮细胞的体外培养体系。Culture system of bovine mammary epithelial celI（BMEC） from milk was established to study the proliferation and differentiation and molecular mechanism of gene express in mammary epithelial cell. The basic medium is DMEM/F12（V/V= 1 ： 1） and 20% fetal bovine serum （FBS） , and insulin（5 μg/mL）, transferring （ 5 μg/mL, hydrocortisone （ 1 μg/mL ）, progesterone （ 1 μg/mL ）, penicillin （ 100 IU/mL ） and streptomycin（ 100 IU/mL） were added. The cell cultured on the plastic petri dishes was covered with collagen. The major morphology of epithelial cells appear to be characteristic guboidal and cobblestone. The phase contrast photomicrographs indicate the presence of domelike and nipple structures in the proliferated cells with the phenomenon of lactescence. The cell biological properties were observed and investigated from the cell planting survival efficiency, cell population doubling time and cell growth curve. The results showed that the BMEC grows well and proliferates vigorously in this culture system. The SDS - PAGE and amplification of （αs1 - casein gene in cells showed that the cultured cell is BMEC, and it has a function that expresses the bovine casein.

关 键 词：乳腺上皮细胞 体外培养体系 奶牛 

分 类 号：S852.1[农业科学—基础兽医学] S823.91[农业科学—兽医学]