人乳腺上皮细胞抽提物重编程人脂肪来源干细胞的实验研究  

Experimental research of reprogramming of adipose-derived stem cells using human mammary epithelial cell extracts

作　　者：杨艳清[1] 江昌艳 张洁[1] 许其军[1] 王海燕 

机构地区：[1]武汉大学附属同仁医院(武汉市第三医院)整形外科,湖北武汉430060 [2]武汉市医学会,湖北武汉430000

出　　处：《中国美容整形外科杂志》2017年第12期709-711,共3页Chinese Journal of Aesthetic and Plastic Surgery

基　　金：湖北省卫生计生科研项目(WJ2015MB145)

摘　　要：目的观察人乳腺上皮细胞抽提物重编程人脂肪来源干细胞(h ADSCs),使其向上皮细胞转化的可行性。方法采用胶原酶消化法分离并获得脂肪来源干细胞;将超声乳化人乳腺上皮细胞离心后获得抽提物冻存备用;培养第3代h ADSCs,链球菌溶血素O(SLO)处理后,实验组6例加入乳腺上皮细胞抽提液,对照组6例不加乳腺上皮细胞抽提液。细胞培养过程中观察h ADSCs的形态学变化,采用蛋白质印迹法检测卵透明带蛋白1(ZO1)和细胞角蛋白18(CK18)表达的变化。结果流式细胞仪检测结果显示,人脂肪组织分离培养的干细胞表达CD13、CD29、CD44、CD90和CD105呈阳性表达,而不表达CD45。实验组细胞重编程后第16天,细胞形态开始转变为上皮样的圆形;第18天蛋白印迹法检测上皮细胞抗原标志物ZO1和CK18表达增加,与对照组比较,差异有统计学意义(P<0.05)。结论人乳腺上皮细胞抽提物可以重编程脂肪来源干细胞,使其向上皮细胞转化。Objective To explore the feasibility of human mammary epithelial cell extracts in the reprogramming and conversion of human adipose-derived stem cells（h ADSCs） to epithelial cells. Methods The ADSCs were isolated by the collagenase digestion method and cultured. The human mammary epithelial cells were obtained by ultrasonic emulsification. The extracts were acquired through high speed centrifugation and frozen for further use. The third generation of h ADSCs were cultured and treated with streptolysin O（SLO）.The mammary epithelial cell extracts were added into the 6 cases in the experimental group, but not added into the 6 cases in the control group. The morphological changes of ADSCs were observed during the cell culture process. The changes in zona occludens protein 1（ZO1）and cytokeratin 18（CK18） expression in each group were detected by Western blot. Results Flow cytometry showed that the expression of CD13, CD29, CD44, CD90, and CD105 were positive and the expression of CD45 was negative. At 16 days after ADSCs reprogramming in the experimental group, the cell morphology began to turn into an epithelial-like circle shape. Western blot results demonstrated that the expression of ZO1 and CK18 in the experimental group increased more significantly than that in the control group at the 18 th day（P0.05）. Conclusion Human mammary epithelial cell extracts can reprogram ADSCs and cause them to transform into epithelial cells.

关 键 词：乳腺上皮细胞 脂肪来源干细胞 重编程 细胞抽提物 

分 类 号：R622[医药卫生—整形外科]