人骨髓间充质干细胞向汗腺样细胞诱导分化的研究  

Study on induction of human bone marrow mesenchymal stem cells into human sweat gland-like cells

作　　者：陈艳[1,2] 赵洪良[2] 谭志军[2] 赵焕军[2] 张翠萍[2] 付小兵[2] 

机构地区：[1]北京军区总医院药理科,北京100700 [2]解放军总医院第一附属医院全军创伤修复与组织再生重点实验室,北京100048

出　　处：《感染．炎症．修复》2014年第2期76-78,F0002,共4页Infection Inflammation Repair

基　　金：国家973计划资助项目(2012CB518105);国家自然科学基金项目(81121004;81230041;81171798)

摘　　要：目的:观察体外热休克汗腺细胞(SGCs)和人骨髓间充质干细胞(BM-MSCs)共培养体系中BMMSCs的形态和表型变化,为进一步表观遗传学表达谱的检测及汗腺诱导关键转录因子的研究提供实验基础。方法:体外分离、培养、扩增人BM-MSCs和SGCs,成骨和成脂诱导分化以鉴定BM—MSCs的分化功能。在Transwell间接共培养体系中,培养的BM-MSCs和经47℃高温处理造成热休克的SGCs在Transwell板中间接共培养;在Transwell+诱导因子共培养体系中,上室的BM-MSCs培养基中添加了汗腺诱导因子(无汗性外胚叶发育不良蛋白、重组人表皮生长因子和胰岛素-转铁蛋白-亚硒酸钠)。监测共培养过程中BM-MSCs的细胞形态变化,免疫荧光法检测诱导后BM-MSCs的表型改变。结果:经与热休克SGCs共培养诱导10 d后,部分BM-MSCs有由长梭形变为扁平状多边形的趋势,且局部细胞间连接紧密成片。BM-MSCs诱导前不表达CEA和CK19;BMMSCs诱导后,Transwell间接共培养体系部分细胞CEA和CKl9表达阳性,Transwell+诱导因子共培养体系CEA和CK19阳性细胞数明显多于Transwell间接共培养体系。结论:热休克汗腺细胞与BM-MSCs在Transwell间接培养以及相关汗腺诱导因子的共培养体系下,BM-MSCs呈现向SGCs诱导分化趋势。Objective：To observe the changes in morphology and phenotype of human bone marrow mesenchymal stem cells （BM-MSCs）after co-cultured with human sweat gland cells （SGCs）,in order to provide the experimental basis for the study on sweat gland cells induction-associated key transcription factors. Methods：BM-MSCs and SGCs were isolated,cultured, expanded and identified with the methods of differentiation to adipocytes and osteocytes invitro respectively. In transwell co-cultured experiment,BM-MSCs were induced into SGCs by indirect co-culture with 47 ℃heat-shocked confluent SGCs using transwell plate.But in transwell＋inducing factors co-culture experiment,BM-MSCs culture medium was supplemented with series of inducing factors,including EDA-A1 ,recombinant human epidermal growth factor and ITS. After being co-cultured, morphological and phenotypic changes in BM-MSCs were observed. The antigen expressions of BM-MSCs were detected by immunofluorescence. Results：After co-cultured with heat-shock SGCs for 10 days,the shape of a part of MSCs changed from long-clostridial shape to flat and polygonal. Non-induced BM-MSCs were negative for CEA and CK1 9. Induced BM-MSCs were positive for CEA and CK19,and the positive cells of transwell ＋ inducing factors co-culture was more abundant than that of transwell co-culture. Conclusions：MSCs could differentiate into sweat gland-like cells at phenotype and shape by a proper in vitro co-culture system of BM-MSCs and SGCs with or without inducing factors,and it was more efficient when they were co-cultured with inducing factors.

关 键 词：骨髓间充质干细胞 汗腺 共培养 表型转化 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]