奶山羊乳腺上皮细胞与骨髓间充质干细胞混合培养下最适共培养体系的建立

Establishment of the Optimal Co-culture System of Dairy Goat Mammary Epithelial Cells and Bone Marrow Mesenchymal Stem Cells in Mixed Culture

【目的】研究乳腺上皮细胞与骨髓间充质干细胞混合培养促进细胞增殖作用的机制,提高奶山羊乳腺上皮增殖水平。【方法】采用分离培养的1月龄健康萨能奶山羊乳腺上皮细胞和骨髓间充质干细胞,分别设乳腺上皮细胞单纯培养组和不同比例的乳腺上皮细胞与骨髓间充质干细胞混合培养组,培养24、48、72、96、120和168 h时观察各组细胞的生长状态并检测细胞增殖率和贴壁率。【结果】混合培养组增值率和贴壁率比单纯培养组高,且细胞扁平时间晚于单纯培养组,混合培养体系可促进BMEC的增殖,减缓细胞凋亡。当BMEC与BMSCs以2∶1的比例混合培养到96 h时细胞的增值率和贴壁率最高。【结论】奶山羊乳腺上皮细胞和骨髓间充质干细胞共培养体系能够减缓乳腺上皮细胞的凋亡,促进乳腺上皮细胞增殖,提高细胞贴壁率。最佳共培养浓度为2∶1,最佳作用时间为96 h。

[ Objective ] To explore the effects of mammary epithelial cells co - cultured with mesenchymal stem cells on cell proliferation. [ Method ] 1 - month - old Saanen dairy goat mammary epithelial cell and bone marrow mesenchymal stem cells were used and mammary epithelial cells and bone marrow mesenchymal stem cells mixed culture group were set up respectively in the simple culture group along with different proportions of mammary epithelial cells. After they were cultured 24 h, 48 h, 72 h, 96 h, 120 h, 168 h, the growth state of the cells was observed and the cell proliferation rate and adherence rate were detected. [ Result] The results showed that appreciation and attachment rate of mixed culture group were higher than those of the simple cul- ture group, and cell flat time was later than pure culture group. Mixed culture system could promote the prolif- eration of BMEC and slow apoptosis. When the ratio of BMEC and BMSCs was mixed in the proportion of 2： 1 to 96 h, the rate of increment and adherence rate of the cells was the highest. [ Conclusion ] The co - culture system of dairy goat mammary epithelial cells and bone marrow mesenchymal stem cells can slow down the ap- optosis of mammary epithelial cells, promote breast epithelial cell proliferation, and increase cell adhesion rate. The best co- culture concentration is 2：1 ,and the time is 96 h.