脂肪间充质干细胞的体外诱导和成血管作用  

Induction of adipose-derived mesenchymal stem cells in vitro and angiogenesis

作　　者：张亚[1] 周云[1] 贾立山[1] 翟景梅[1] 苗小芬 王睿[2] 

机构地区：[1]苏州大学附属儿童医院外科,215003 [2]苏州中医院病理科

出　　处：《中华小儿外科杂志》2011年第5期380-383,共4页Chinese Journal of Pediatric Surgery

基　　金：基金项目：江苏省卫生厅面上科研课题项目资助（编号：H200819）,苏州市社会发展科技项目资助（编号：sS08021）

摘　　要：目的 研究在体外诱导脂肪间充质干细胞（ADMSCs）向内皮细胞分化、在立体培养基上的血管形成情况.方法 将传至第三代的ADMSCs用内皮细胞诱导液、Matrigel三维培养基进行诱导培养,对ADMSCs和诱导细胞选用CD31、CD44细胞表面抗原在流式细胞仪检测表达情况,用HE染色、FⅧ-RAg免疫组织化学染色及倒置显微镜、透射电镜等进行观察鉴定.结果流式细胞仪上检测ADMSCs的表达为CD44阳性、CD31阴性,诱导的细胞CD31阳性、CD44阴性.诱导细胞14d倒置显微镜下呈铺路石样形态,FⅧ-RAg染色细胞呈阳性,透射电镜下在细胞浆内见到内皮细胞特有标志物Weibel-Palade小体。ADMSCs在Matrigel三维培养基上诱导,24h细胞迁徙成团、有伪足伸出,诱导7d伸出的细胞形成交叉网格状,13d形成较长血管,20d较长血管粗而多、出现小分叉,30d长血管、分叉血管变粗厚;FⅧ-RAg染色后诱导血管亦呈阳性.结论 脂肪间充质干细胞在体外经诱导能向内皮细胞分化、能形成血管,可作为促进组织工程移植物血管化的良好的种子细胞.Objective To study in vitro induction of adipose-derived mesenchymal stem cells （ADMSCs） into endothelial cells and blood vessel formation on three-dimensional （3D） media. Methods The 3rd passage ADMSCs were induced in vitro into endothelial cells on conditional medium and grew on Matrigel medium. The cell surface antigens CD31 and CD44 were detected with flow cytometric analysis before and after induction. HE staining, FⅧ-RAg immunohistochemical staining, inverted microscope and transmission electron microscope were used for morphological study.Results The expression of CD44 was positive and CD31 negative in ADMSCs in flow cytometric analysis.After induction,CD31 became positive while CD44 was negative. Paving-stone-like cell appearance was seen under inverted microscope 14 days after induction.The cells were FⅧ-RAg positively stained with immunohistological method,and Weibel-Palade body was observed under transmission electron microscope.On Matrigel media,the induced cells migrated in lumping with pseudopodia protrusion after 24 hours,and formed grid structure on the 7th day.Long vasculature was observed on the 13th day,and the vessels branched on the 20th to 30th day.The vessels stained positive for FⅧ-Rag. Conclusions ADMSCs have the potential to give rise to endothelial cells with in vitro induction and to form vessel structure in 3D media.ADMSCs have potential role in vascularized tissue engineering grafting.

关 键 词：脂肪间充质干细胞 内皮细胞 血管 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]