比较不同诱导体系体外诱导华通胶间充质干细胞分化为内皮细胞的研究  

Study of umbilical cord-derived human mesenchymal stem cells differentiation into endothelial cells in different induction conditions in vitro

作　　者：曹芳英[1,2] 张宁坤[2] 高连如[2] 朱智明[2] 

机构地区：[1]南方医科大学研究生部,广东广州510515 [2]海军总医院心脏中心,北京100048

出　　处：《转化医学杂志》2013年第6期324-328,共5页Translational Medicine Journal

基　　金：海军总医院创新培育基金(CX201101)

摘　　要：目的比较诱导因子和非接触共同培养2种方法诱导人脐带华通胶间充质干细胞(human umbilical cord Wharton's jelly-mesenchymal stem cells,HUW-MSCs)分化为内皮细胞的优劣。方法剖宫产取脐带用酶消化和组织块贴壁2种方法获得HUW-MSCs。实验分诱导因子组、共同培养组和单纯培养组。诱导因子组用血管内皮细胞生长因子、碱性成纤维细胞生长因子、表皮生长因子联合诱导培养;共同培养组HUWMSCs与人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)非接触共同培养;单纯培养组只加培养液培养。培养周期为14 d。第7天检测CD309,第10天进行Dil标记乙酰化低密度脂蛋白(Dil-labeled acetylated low density lipoprotein,Dil-Ac-LDL)吞噬实验;第14天检测CD31、血管假性血友病因子(von Willebrand factor,vWF)、内皮素-1(endothelin-1,ET-1)和一氧化氮(nitric oxide,NO)。结果各组细胞形态较实验前有显著变化,但均未见典型铺路石样改变。3组细胞表达CD309上调;诱导因子组和单纯培养组表达CD31下降,而共同培养组CD31明显上调。NO及ET-1含量检测均较HUVECs低,但差异无统计学意义(P>0.05),3组之间比较差异同样无统计学意义(P>0.05)。vWF及Dil-Ac-LDL各组反应呈弱阳性。结论 HUW-MSCs在诱导因子和内皮细胞旁分泌作用下均具有向内皮细胞分化倾向,使用诱导因子较非接触共同培养方法诱导HUW-MSCs向内皮分化的效果更好。Objective To study the difference of umbilical cord-derived human umbilical cord Wharton＇s mesenchymal stem cells （HUW-MSCs） differentiation into endothelial ceils between methods by inducing factors and non-contact co-culture. Methods The umbilical cords were ob- tained from cesarean section. We isolated the Wharton＇s jelly and obtained human umbilical veins endothelial cells （HUVECs）. HUW-MSCs were divided into 3 subgroups, factors group, cocuhure group and haplo-cultivation group. The culture media of induction subgroup contained vascular en- dothelial growth factor （VEGF）, basic fibroblast growth factor （bFGF）and epidermal growth factor （EGF）. MSCs were first cocultured with HUVECs in non-contact system. Media for haplo-cultiva- tion was just MI99. All of the M199 contained 10% FBS and 1% mycillin. The whole cultivation cy- cle was fortnight, and we checked out the CD309 in d7, phagocytosis of Dil-labeled acetylate low density lipoprotein （Dil-Ac-LDL）was done in dlO and experiments of CD31, von Willebrand factor （vWF）, nitric oxide （NO） and endothelin （ ET-I ） were finished in d14. Results Morphous of cell in 3 subgroups had changed dramatically compared to MSCs cultured by DMEM/Ham＇s F12, but none show as endothelial cell. The expression of CD309 showed up-regulation in 3 subgroups. Expression of CI）31 increased in coculture group, but decreased in other two groups. Both content of NO and ET-1 were highest in factors＇ group, coculture group was next. There was no significance among three groups. Conclusion Both factors and paracrine secretion affect HUW-MSCs differentiation into vascular endothelial cells, and the effects of factors are stronger.

关 键 词：脐带华通胶 间充质干细胞 血管内皮细胞 生长因子 非接触共培养 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] Q254[医药卫生—基础医学]