作　　者： (赵艳坤）; (邵伟）; (雒诚龙）; (余雄）;

机构地区： 新疆农业大学动物科学学院,新疆肉乳用草食动物营养实验室,乌鲁木齐830052

出　　处： 《中国实验动物学报》 2017年第4期391-398,共8页

摘　　要： 目的旨在探究无血清共培养条件下脐带间充质干细胞(umbilical cord mesenchymal stem cells,UCMSCs)对奶牛乳腺上皮细胞(bovine mammary gland epithelial cells,BMECs)的增殖作用。方法选取生长状态最佳的UC-MSCs和BMECs,将两种细胞以直接接触和间接接触两种共培养方式作为试验组,直接接触是按照UCMSCs:BMECs分别为2∶1、1∶1、1∶2、1∶3、1∶4、1∶5、1∶10的浓度梯度混合共培养,间接接触则是提取UC-MSCs的上清液作为条件培养基重悬BMECs,对照组为UC-MSCs和BMECs单纯培养组,并设阴性空白对照,于0、4、8、12、24、36、48、60、72 h时观察各组细胞的生长变化,采用CCK-8法检测各组细胞增殖情况。结果在48 h,条件培养基BMECs组A值显著高于对照组(P<0.05),而1∶2浓度组A值达到峰值,极显著高于对照组(P<0.01),显著高于其他各浓度组和条件培养基BMECs组(P<0.05)。结论无血清条件下,UC-MSCs和BMECs共培养可以促进BMECs增殖,直接接触促增殖效果优于间接接触,且最适比例为1∶2,最佳时间是48 h。 Objective To explore the proliferation-promoting effect of bovine mammary gland epithelial cells（ BMECs） co-cultured with umbilical cord mesenchymal stem cells（ UC-MSCs） in serum-free culture mediuum. Methods Bovine UC-MSCs and BMECs were selected for co-culturing in direct or indirect contact. In the direct contact culture groups,UC-MSCs and BMECs were co-cultured at concentration ratios of 2∶ 1,1∶ 1,1∶ 2,1∶ 3,1∶ 4,1∶ 5,and 1： 10,respectively. In the indirect contact culture group,the supernatant of UC-MSCs was used as the conditioned medium to resuspend BMECs. In the control groups,UC-MSCs and BMECs were cultured alone. The cell growth status in each group was observed at 0,4,8,12,24,36,48,60,72 h after culture,and cell proliferation was detected by cell counting kit-8（ CCK-8） assay. Results At 48 h,the optical density of the conditioned medium-BMECs group was significantly higher compared with the control groups（ P〈0. 05）. Meanwhile,the optical density in the direct contact group at a concentration ratio of 1∶ 2 reached the peak,which was extremely significantly higher compared with the control groups（ P〈0. 01） and significantly higher compared with the other direct contact culture groups and the conditioned medium-BMECs group（ P〈0. 05）. Conclusions Co-culture of UC-MSCs and BMECs in serum-free culture medium is capable to promote the proliferation of BMECs,and the co-culture by cell-to-cell contact has a better effect. The optimal concentration ratio of UC-MSCs to BMECs is 1∶ 2,and the optimal culture time is 48 h.

关 键 词： 脐带间充质干细胞 乳腺上皮细胞 无血清 共培养 增殖