人脂肪间充质干细胞诱导为内皮样细胞的可行性及其小鼠眼内应用短期安全性  

The feasibility of human adipose-derived mesenchymal stem cells induced to endothelial-like cells and the safety of induced cells intraocular application in a short period

作　　者：郭凯[1] 罗燕[2] 李士清[3] 田景毅 Guo Kai;Luo Yan;Li Shiqing;Tian Jingyi(Department of Ophthalmology,The Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010059,China;Zhongshan Ophthalmic Center,Sun Yat-sen University,Guangzhou 510060,China;Henan Eye Institute,Henan Eye Hospital,Henan Provincial People's Hospital,Zhengzhou 450003,China)

机构地区：[1]内蒙古医科大学附属医院眼科,呼和浩特010059 [2]中山大学中山眼科中心,广州510060 [3]河南省人民医院,河南省立眼科医院,河南省眼科研究所,郑州450003

出　　处：《中华实验眼科杂志》2019年第11期880-883,共4页Chinese Journal Of Experimental Ophthalmology

基　　金：国家自然科学基金项目(81760173);内蒙古自治区自然科学基金项目(2019MS08014)。

摘　　要：目的评估人脂肪间充质干细胞(hADSCs)向内皮样细胞诱导分化的可行性及其眼内应用的安全性。方法体外培养并诱导hADSCs向内皮样细胞分化,实验组加入青链霉素、血管内皮生长因子(VEGF)、碱性成纤维细胞生长因子(bFGF),对照组分别加入等量磷酸盐缓冲液(PBS),分别观察内皮细胞标志物血管性血友病因子(vWF)的表达。应用随机数字表法将6只C57BL/6J小鼠分为实验组和对照组,每组3只,实验组小鼠玻璃体腔注射诱导细胞,对照组注射等量PBS,电子显微镜下分别观察小鼠视网膜细胞变化,评估诱导后细胞眼内应用的安全性。结果hADSCs体外诱导后表达内皮细胞标志物vWF,实验组vWF免疫荧光呈绿色强荧光,显色率为100%,正常对照组vWF免疫荧光无显色,显色阳性率为0。诱导细胞小鼠玻璃体腔注射后1个月视网膜神经节细胞及视杆细胞未见变性、坏死等毒性改变。结论hADSCs体外诱导可以向内皮样细胞分化,小鼠眼内短期应用无视网膜毒性反应。Objective To evaluate the feasibility of differentiation of human adipose mesenchymal stem cells(hADSCs)into endothelial-like cells and the safety of induced cells intraocular application.Methods HADSCs were induced to endothelial-like cells in vitro.The experimental group was added with streptomycin,vascular endothelial growth factor(VEGF)and basic fibroblast growth factor(bFGF),and the control group was added with the same amount of phosphate buffered saline(PBS),the expression of von Willebrand factor(vWF),a marker of endothelial cells,was observed in the two groups.Six C57BL/6J mice were randomly divided into experimental group and control group by using the random number method,3 mice for each group,the induced cells were injected intravitreally into C57BL/6J mice as experimental group,and the same amount of PBS buffer was injected intravitreally into C57BL/6J mice as control group.The changes of retinal cells were observed by electron microscopy.The use and care of the animals complied with Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission.Results After induction of hADSCs in vitro,the endothelial cell marker was expressed.VWF immunofluorescence of the experimental group showed strong green fluorescence,and the color rendering rate was 100%.The control group showed no coloration of vWF immunofluorescence,and the color positive rate was 0.After 1 month of intravitreal injection,the retinal ganglion cells and rod cells were not degraded and necrotic.Conclusions HADSCs can differentiate into endothelial-like cells in vitro,and there is no retinal toxicity in a short-term.

关 键 词：脂肪间充质干细胞 内皮样细胞 诱导分化 毒性反应 

分 类 号：R73[医药卫生—肿瘤]