人诱导性多能干细胞分化的血管平滑肌细胞体外培养圆环形工程化组织的实验研究  

Experimental study of culturing annular engineering tissue in vitro by vascular smooth muscle cells de⁃rived from human induced pluripotent stem cells

作　　者：周嘉辉[1,2] 马文韬 姚博谦 罗玮 方丽君 刘鹏 林展翼[1,2,3] ZHOU Jia-hui;MA Wen-tao;YAO Bo-qian;LUO Wei;FANG Li-jun;LIU Peng;LIN Zhan-yi(Guangdong Cardiovascular Institute,Guangdong Provincial People′s Hospital,Guangdong Academy of Medi-cal Sciences,Guangzhou 510080,China;Guangdong Provincial People′s Hospital,Guangdong Academy of Medical Sciences,Guangzhou 510080,China;School of Medicine,South China University of Technology,Guangzhou 510006,China;The Third People′s Hospital of Dongguan,Dongguan,Guangdong 523326,Chi-na;Nanfang Hospital of Southern Medical University,Guangzhou 510515,China;School of Biology and Biological Engineering,South China University of Technology,Guangzhou 510006,China)

机构地区：[1]广东省心血管病研究所,广东省人民医院(广东省医学科学院),广州510080 [2]广东省人民医院(广东省医学科学院),广州510080 [3]华南理工大学医学院,广州510006 [4]广东省东莞市松山湖中心医院,广东东莞523326 [5]南方医科大学南方医院,广州510515 [6]华南理工大学生物科学与工程学院,广州510006

出　　处：《岭南心血管病杂志》2021年第6期736-739,共4页South China Journal of Cardiovascular Diseases

基　　金：广东省人民医院“登峰计划”科研专项(项目编号:DFJH201904)。

摘　　要：目的通过体外培养圆环形工程化组织,建立一种评价人诱导性多能干细胞(induced pluripotent stem cells,iPSC)来源的种子细胞应用于组织工程培养的实验方法。方法采用拟胚体法诱导人iPSC分化为血管平滑肌细胞,经免疫荧光染色确认细胞类型,把种子细胞加入琼脂基质培养得到圆环形工程化组织,行组织切片染色评价种子细胞的增殖和分泌功能。结果用拟胚体法诱导人iPSC可获得大量表达血管平滑肌细胞特异标志物的种子细胞。在体外把该细胞加入琼脂基质中培养获得圆环形工程化组织,组织的形态规整,具有一定的力学强度,能自我维持组织的三维形态;内部含有以胶原蛋白为主的细胞外基质,基质间细胞排列紧密,持续表达成熟的血管平滑肌细胞标志物。结论人iPSC分化的血管平滑肌细胞在体外被成功培养出结构完整的圆环形工程化组织,对组织的形态、结构及细胞外基质分析的结果显示,其可作为人iPSC来源的种子细胞应用于组织工程培养的评价指标。Objectives To establish an experimental method for evaluating the application of seed cells derived from human induced pluripotent stem cells(iPSC)in tissue engineering via cultivating annular tissue in vitro.Methods Human iPSC was induced to differentiate into vascular smooth muscle cells(VSMCs)by embryoid body method.After confirmed by immunofluorescence staining,the induced VSMCs were added to an agar matrix mould to cultivate to be an annular engineering tissue.Later the amount of seed cells and extracellular matrix were evaluated by histological staining.Results A large number of seed cells expressing VSMCs specific markers were obtained by inducing human iPSC with embryoid body method.Cells were cultured in agar mould for 7 days to form an annular engineering tissue in vitro,which had regular appearance,certain mechanical strength and self-sustaining tissue morphology.There was collagen based extracellular matrix into the tissue.Seed cells distributed closely among the extracellular matrix and continuously expressed mature VSMCs markers.Conclusions iPSC-derived VSMCs were successfully cultured into annular tissue with complete structure in vitro.The morphology,structure and extracellular matrix of the tissue can be deserved as evaluations for the application of seed cells derived from human iPSC in tissue engineering.

关 键 词：血管平滑肌细胞 组织工程 种子细胞 诱导性多能干细胞 

分 类 号：R33[医药卫生—人体生理学]