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背景:乳腺干细胞的培养对研究乳腺的发育及乳腺癌具有重要的意义。目的:找到一种易操作的体外分离培养乳腺干细胞的方法,并验证其安全性。方法:分离培养乳腺癌旁正常组织中乳腺上皮细胞,流式细胞仪分选CD49f和EPCAM阳性细胞进行干细胞表面标记分析及细胞集落形成能力分析,然后在乳腺干细胞、乳腺癌组织和乳腺癌旁正常组织中提取mR NA,实时荧光定量PCR检测C-erbB-2和Maspin基因的表达。结果与结论:(1)成功培养出人乳腺干细胞,并且细胞表面特异性标志物CD49f,EPCAM高表达;(2)集落形成能力分析可见混合集落、腔上皮细胞集落、肌上皮细胞集落;(3)乳腺干细胞C-erbB-2低表达,而Maspin高表达,证明来源于乳腺癌正常组织的乳腺干细胞具有生物安全性。
BACKGROUND: Breast stem cell culture is of great importance for studying the development of mammary gland and breast cancer. Objective: To find an easy-to-manipulate method for the isolation and culture of breast stem cells in vitro and to verify its safety. Methods: Breast epithelial cells were isolated and cultured in normal tissue adjacent to breast cancer. CD49f and EPCAM positive cells were sorted by flow cytometry for stem cell surface marker analysis and colony forming ability analysis. Then normal breast cancer cells, breast cancer tissues and normal breast tissues Tissues extracted mR NA, real-time fluorescence quantitative PCR detection of C-erbB-2 and Maspin gene expression. RESULTS AND CONCLUSION: (1) Human breast stem cells were successfully cultured, and the cell surface markers CD49f and EPCAM were highly expressed. (2) The colonies of colony, luminal cell colonies and myoepithelial cell colonies were observed. (3) ) Breast cancer stem cell C-erbB-2 low expression, Maspin high expression, proof of breast cancer stem cells derived from normal tissue biosafety.