PKH26标记人脂肪间充质干细胞及眼内示踪的可行性研究  

The feasibility of PKH26-marked human adipose derived mesenchymal stem cells and intraocular trace

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876-879,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PKH26对人脂肪间充质干细胞(hADSCs)进行体外标记及眼内标记的可行性。方法体外培养hADSCs,使用PKH26对hADSCs进行体外标记并观察标记结果。采用抽签法将C57BL/6J小鼠18只分为正常对照组(6只)、标记细胞组(9只)和未标记细胞组(3只),标记细胞组和未标记细胞组C57BL/6J小鼠玻璃体腔分别注射2×10^5/ml标记细胞和未标记细胞各1μl,注射后1个月分别行视网膜铺片观察荧光标记结果。取正常对照组及标记细胞组小鼠视网膜进行苏木精-伊红染色及电子显微镜检查判定毒性反应。结果hADSCs经PKH26体外标记后细胞膜呈红色荧光,标记细胞组小鼠玻璃体腔注射标记细胞后1个月可见视网膜血管前红色强荧光,标记细胞组小鼠视网膜苏木精-伊红染色及电子显微镜检查均未见细胞变性坏死及增生。hADSCs体外、体内标记显影结果显示,未标记细胞组均无荧光显色,显色阳性率均为0,标记细胞组均出现红色荧光,显色阳性率均为100%。结论PKH26可用于标记hADSCs及眼内示踪,形态学评估显示其无毒性反应。Objective To evaluate the feasibility of PKH26 marked human adipose derived mesenchymal stem cells(hADSCs)in vitro and intraocular.Methods HADSCs were cultured in vitro and marked with PKH26.Eighteen C57BL/6J mice were divided into normal control group(6 mice),labeled cell group(9 mice)and unlabeled cell group(3 mice)by using sortition randomization method,labeled cells were injected intravitreally in C57BL/6J mice as labeled cell group,and the unlabeled cells were injected intravitreally in C57BL/6J mice as unlabeled cell group.After 1 month,retinal slab was checked to contrast the results of intraocular labeling.The retina was taken out for hematoxylin-eosin staining and electron microscopy to observe the toxicity.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 In vitro,red fluorescent was found in the cytomembranes after hADSCs were labeled by PKH26.Retinal patch of labeled cell group showed red fluorescence of hADSCs were in front of the retina.Hematoxylin-eosin staining of retinal tissue and optic ganglion cells showed that cell degeneration and proliferation were not found in labeled cell group.The hADSCs in vivo and in vitro marked enhancement results showed that,no fluorescent was found in the unlabeled cell group,the color positive rate were both 0,while red fluorescence was found in the labeled cell group,the color positive rate were both 100%.Conclusions PKH26 can be used to mark and intraocular trace of hADSCs.Also,it has no morphology toxic reaction.

关 键 词：脂肪间充质干细胞 PKH26 细胞标记 毒性反应 

分 类 号：R73[医药卫生—肿瘤]