体外构建干细胞汗腺细胞融合体的初步研究  

Preliminary study on the establishment of fusion body of human umbilical cord mesenchymal stem cells with human sweat gland cells in vitro

作　　者：刘坤坤 赵亚平[1] 高玮[2] 高慧[1] 谢淼[1] LIU Kunkun;ZHAO Yaping;GAO Wei(Department of Plastic Surgery,The First Hospital of Shijiazhuang City,Hebei,Shijiazhuang 050011,China)

机构地区：[1]河北省石家庄市第一医院整形科,050011 [2]河北省石家庄市第一医院预防保健处,050011

出　　处：《河北医药》2019年第20期3130-3133,共4页Hebei Medical Journal

摘　　要：目的研究人脐带间充质干细胞(UCMSC)与人汗腺细胞(SGC)形成的合核细胞生物学特性,为实现汗腺再生探索新的方法和途径。方法体外分离培养UCMSC和SGC,通过检测CD14、CD44、CD29、CD34、CD105、CD45的表达情况来鉴定UCMSC,检测CK19、CEA的表达情况来鉴定SGC。制取并检测间充质干细胞/汗腺细胞合核细胞,用PKH26、CFSE分别对UCMSC、SGC进行标记,通过聚乙二醇(PEG)化学融合法制备间充质干细胞/汗腺细胞融合体,用荧光显微镜荧光观察并筛选出融合细胞后进行培养;1周后用流式细胞仪检测合核细胞中CD14、CD44、CD29、CD34、CD105、CD45表达率,用免疫组织化学检测CK19、CEA表达情况。结果间充质干细胞/汗腺细胞合核细胞的免疫组化检测CK19和CEA表面抗原均为阳性,流式细胞术检测表面标志物CD29、CD44、CD105表达率较高。结论诱导融合后的间充质干细胞/汗腺细胞合核细胞同时具备干细胞和汗腺细胞的表型。Objective To investigate the biological characteristics of fusion cells of human umbilical cord mesenchymal stem cells(MSCs)with human sweat gland cells sweat gland cells(SGCs)in vitro,in order to explore an new way to realize sweat glands regeneration.Methods The MSCs and SGCs were isolated,cultured and amplified in vitro,then were stained by PKH26 and CFDA SE,respectively.The UCMSCs were identified by detecting the expression levels of CD14,CD44,CD29,CD34,CD105,CD45,and the SGCs were identified by detecting the expression levels of CK19 and CEA.The fusion cells of MSCs and SGCs were made by polyethylene glycol(PEG),which were were marked by PKH26 and CFSE,respectively.The fusion cells were screened by fluorescence microscopy,then were cultured in vitrol.After 1 week,the expression levels of CD14,CD44,CD29,CD34,CD105,CD45s in fusion cells were detected by flow cytometry,and the expression levels of cytokeratin 19(CK19)and carcinoembryonic antigen(CEA)in fusion cells were detected by immunohistochemical method.Results The surface antigens of CK19 and CEA in fusion cells of MSCs and SGCs were positive.The results of flow cytometry showed that the expression levels of CD29,CD44,CD105 in fusion cells of MSCs and SGCs were higher,however,the expression levels of CD29 and CD44 were relatively lower.Conclusion The fusion cells of MSCs and SGCs have simultaneously the biological characteristics of mesenchymal stem cells and sweat gland cells.

关 键 词：间充质干细胞 汗腺细胞 融合细胞 再生 聚乙二醇化学融合法 

分 类 号：R758.74[医药卫生—皮肤病学与性病学]