牛乳腺上皮细胞体外培养条件的优化及功能验证  

Optimization of Culture Condition and Functional Verification for Bovine Mammary Epithelial Cells in Vitro

作　　者：贺小英[1] 马利兵[1] 程腾[1] 刘希宇 刘娟[1] 吴飞[1] 

机构地区：[1]内蒙古科技大学数理与生物工程学院,中国内蒙古包头014010

出　　处：《生命科学研究》2015年第1期19-23,共5页Life Science Research

基　　金：国家自然科学基金项目(31160245);内蒙古自然科学基金项目(2012MS0503);内蒙古自治区高等学校青年科技英才支持计划资助项目

摘　　要：乳腺上皮细胞是研究泌乳的调控机理、乳腺癌发病机制以及制备乳腺生物反应器靶细胞。由于乳腺细胞体外培养生命周期较短并且增殖活力与体外培养时间呈负相关,到目前为止,能够适合体外研究的乳腺细胞系报道较少。因此,在优化了乳腺细胞体外培养条件的基础上,进一步验证了优化体系后培养的乳腺上皮细胞生物学的稳定性、蛋白表达情况及转基因的效率。结果表明:1∶1 DMEM/F12基础培养液+10%胎牛血清+2 mmol/L谷氨酰胺+10 ng/m L表皮生长因子+10 ng/m L碱性成纤维细胞生长因子+10μg/m L的ITS+5μg/m L胰岛素+0.1 mmol/L非必须氨基酸是维持乳腺上皮细胞体外培养的最适条件。在此培养条件获得的乳腺上皮细胞生长旺盛,传代次数能延长到20代,生长后期仍然能稳定表达CK18,且具有较高的转基因效率。Mammary epithelia cells are an important in vitro model to study the mechanism of lactation,cancer and mammary bioreactor. However, it is very hard to culture in vitro. Currently, fewer cell lines have been used in mammary research field. A valid method for the study of the mammary gland epithelium cell was established. The results showed that the media added 1∶1 DMEM/F12, plus 10% FBS, 2 mmol/L L-glutamine, 10 ng/mL EGF, 10 ng/mL b FGF, 10 μg/m L ITS, 5 μg/m L insulin and 0.1 mmol/L NAEE was the suitable media for bovine mammary epithelial cells. The mammary epithelia cells obtained under optimized condition grew vigorous, extended life span to 20 passages, steadily expressed CK18 and had high transgenic efficiency.

关 键 词：乳腺细胞 体外培养 转基因 乳腺生物反应器 

分 类 号：Q2-0[生物学—细胞生物学]