人脐带间充质干细胞源胞外囊泡促进施万细胞增殖和迁移

Human umbilical cord mesenchymal stem cells-derived extracellular vesicles promote proliferation and migration of schwann cells

目的:探讨人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,hUCMSCs)源胞外囊泡(extracellular vesicles,EVs)对施万细胞增殖和迁移的影响。方法:应用超高速离心法从hUCMSCs上清液中分离和纯化EVs(hUCMSC-EVs),透射电镜观察EVs形态特征;蛋白质印迹法检测EVs标志性蛋白CD63和CD9的表达;通过CCK-8和Transwell小室实验检测不同浓度的hUCMSC-EVs对施万细胞增殖和迁移能力的影响;共聚焦显微镜观察原代施万细胞对EVs的内吞。结果:hUCMSC-EVs呈圆形或椭圆形,且表达特异性分子标志物CD9和CD63;与对照组相比,hUCMSC-EVs显著促进施万细胞增殖和迁移(P均<0.05);hUCMSC-EVs与施万细胞孵育后细胞膜融合。结论:hUCMSC-EVs可显著促进施万细胞的增殖和迁移且呈浓度依赖。

Objective:To investigate the effect of extracellular vesicles(EVs)derived from human umbilical cord mesenchymal stem cells(hUCMSCs)on proliferation and migration of schwann cells.Methods:EVs was isolated and purified from supernatant of hUCMSCs by ultra-high speed centrifugation,morphological characteristics of EVs were observed by transmission electron microscope.Western blotting was used to detect the expression of EVs marker proteins CD63 and CD9.The effects of different concentrations of hUCMSC-EVs on proliferation and migration of schwann cells were tested by CCK-8 and Transwell chamber experiments.Immunofluorescence assay was used to observe the fusion of EVs and primary schwann cells cell membranes.Results:hUCMSC-EVs was round or oval and expressed specific molecular markers CD63 and CD9;Compared with the control group,hUCMSC-EVs significantly promoted schwann cells proliferation(P<0.05)and migration(P<0.05).hUCMSC-EVs fuse with cell membrane after incubation with schwann cells.Conclusion:hUCMSC-EVs can significantly promote proliferation and migration of schwann cells in a concentration-dependent manner.