两种不同消化法对小鼠乳腺上皮细胞培养的影响  

Effects of Two Different Digestion Methods on Culture of Mouse Mammary Epithelial Cells

作　　者：杨航[1,2] 易琼[1,2] 李圆方[1,2] 张小艺[1,2] 王鲁[2] 

机构地区：[1]贵州大学动物科学学院,贵州贵阳550025 [2]贵州省生化工程中心,贵州贵阳550025

出　　处：《动物医学进展》2014年第3期59-64,共6页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(31060347)

摘　　要：为了探究体外培养小鼠乳腺上皮细胞的消化方法,体外分离小鼠乳腺组织,分别采用胶原酶和胰蛋白酶混合消化法或两步消化法消化小鼠乳腺组织,并进行细胞培养,利用角蛋白18激光共聚焦显微技术鉴定目的细胞。结果显示,混合酶消化法能在短期内分离到大量的乳腺上皮细胞,且细胞纯度达到98%,细胞可连续培养至第3代;两步酶消化法虽能分离到一定量的乳腺上皮细胞,但细胞继代培养困难,且细胞不易纯化。以上结果表明,混合酶消化法培养小鼠乳腺上皮细胞优于两步酶消化法。In order to explore the enzyme digestion on mouse mammary epithelial cells（MECs）cultureed in vitro,the mammary tissue was isolated from mice aseptically in vitro,collagenase/trypsin digestion and two-step enzyme digestion were used to culture mouse MECs.The keratin 18,a specific marker for epithe-lial cell,was identifiedby confocal lasers scanning microscope.The results showed that plenty of pure mouse MECs can be harvested by using collagenase/trypsin digestion and continuously cultured for 3 generations. A certain amount of cells can be harvested with two-step enzyme digestion,but cells were subcultured dif-ficultly,and hard to purify.These results suggest that collagenase/trypsin digestion can be preferably used to culture mouse MECs.

关 键 词：小鼠乳腺上皮细胞 胶原酶 胰蛋白酶 混合消化 两步消化 

分 类 号：Q813.11[生物学—生物工程]