体外诱导乳房内脂肪来源干细胞向上皮细胞分化的实验研究

杨杰，郭能强，孙家明等.体外诱导乳房内脂肪来源干细胞向上皮细胞分化的实验研究[J].中华整形外科杂志.2014, 30(3):210-214.

脂肪干细胞;细胞分化;乳腺，人

杨杰，郭能强，孙家明，熊凌云，王荣荣

华中科技大学同济医学院附属协和医院整形外科,武汉,430022

2014年

210-214

万方

目的 观察在体外Transwell系统中共培养后,乳房脂肪来源干细胞(human breast adipose-derived stem cells,hbASCs)向乳腺上皮样细胞转化的能力.方法 自乳房缩小整形术切除的腺体内的脂肪组织中获取hbASCs,取HBL-100细胞系和第3代hbASCs,按1∶1的细胞比例分别接种于Transwell共培养系统的上室和下室,同时设置相同浓度单纯第3代hbASCs为对照组,分别在DMEM/F12培养基中培养,于相差显微镜下观察实验和对照组下室细胞形态变化及生长情况,透射电子显微镜下观察下室细胞超微结构的改变,并对细胞爬片进行荧光免疫细胞化学染色,以鉴定乳腺上皮细胞特异性标志物CK18、CK19的表达.结果 Transwell体系中各细胞均可良好贴壁生长.共培养至15 d,实验组下室细胞数量较对照组明显增多,体积变小,排列呈铺路石样,部分细胞形态上表现出乳腺上皮样细胞特征;电镜下可见微绒毛、桥粒和张力丝等典型上皮细胞结构特征;免疫荧光结果显示,经共培养诱导后,实验组部分细胞可阳性表达乳腺上皮细胞特异性标记物CK18、CK19;对照组诱导细胞上述检测结果均为阴性.实验组CK18、CK19阳性率分别为(24.4±12.0)％和(21.6±16.4)％;对照组分别为(1.8±1.7)％和(1.1±0.6)％,2组比较差异有统计学意义(P＜0.01).结论 与HBL-100细胞共培养后,乳房脂肪干细胞可以被成功诱导成乳腺上皮样细胞. Objective To explore the feasibility of the transdiferentiation of human breast adiposederived stem cells (hbASCs) into mammary epithelial-like cells after co-culturing in Transwell in vitro.Methods The third passage hbASC and the HBL-100 cell line were co-cultured in a Transwell culture system for 15 days.The hbASCs were observed and identified by invcrted phase contrast microscope and transmission electron microscopy,and immunocytochemistry staining in the induced and control groups.Results Both the third passage hbASCs and the HBL-100 cell line cells could adhere and grow rapidly after co-culture in the Transwell system.After co-culture for 15 days,the morphology of some induced hbASCs changed into epithelial-like cells.Some induced hbASCs showed positive expression of CK18,CK19 by immunocytochemistry staining,and typical epithelium cells with microvilli,desmosomes and tonofilaments observed under TEM.The positive rate of CK18 and CK19 was (24.4 ± 12.0)％ and (21.6 ± 16.4)％ in experimental group,and(1.8 ± 1.7)％ and(1.1 ±0.6)％ in control group.Conclusion The data suggests that hbASCs may have the potential to transdifferentiate into human mammary epithelial-like cells after co-culturing in Transwell in vitro.。