内皮细胞外泌体对神经干细胞活力的影响研究

Effects of Endothelial Cell Exosomes on the Vitality of Neural Stem Cells

目的探讨鼠微血管内皮细胞(bEnd.3)外泌体对神经干细胞活力的影响。方法大量培养鼠微血管内皮细胞(bEnd.3),收集细胞上清后提取外泌体并予以鉴定。对神经干细胞的作用分为外泌体组和对照组,其中外泌体组为含有60μg/mL外泌体的培养基培养,对照组为无外泌体培养基培养。采用Transwell共培养72h后检测bEnd.3对神经干细胞的细胞活力影响,采用CCK8法检测对照组和外泌体组细胞培养72 h后的细胞活力,采用平板细胞克隆实验检测2组细胞的增殖能力。结果与对照组比较,Transwell共培养72h后细胞的活力有所提高(1.16±0.52)%vs(3.99±0.12)%。与对照组比较,CCK8法检测外泌体组细胞培养72h时细胞活力明显提高(0.13±0.02)vs(0.18±0.02),细胞平板克隆实验检测结果为(5.6±1.52)vs(13.2±1.92),差异均有统计学意义(P<0.05)。结论鼠微血管内皮细胞(bEnd.3)外泌体可影响神经干细胞的细胞活力。

Objective To investigate the effect of exosomes of bEnd. 3 cells on the vitality of neural stem cells. Methods A large number of bEnd. 3 cells were cultured, and the exosomes were extracted from the supernatants and were identified;there were 2 groups： exosome group（cultured in the medium containing 60 ug/ml exosomes） and control group（cultured in the medium without exosomes） The effect of bEnd. 3 on the cell vitality of neural stem cells was detected after transwell coculture for 72 h. The cell vitality in the 2 groups after 72 h culture was detected by cell counting kit-8（CCK-8） and the cell proliferation was detected by plate cell clone assay. Results After transwell co-culture for 72 hours, the cell vitality in exosome group increased much more than that in control group [（3.99±0.12） % vs（1.16±0.52） %; CCK8 detection indicated a greater increase of cell vitality in exosomes group（0.18±0.02） than in control group（0.13±0.02） while the plate cell clone assay showed out the same result [exosomes group vs control group：（13.2±1.92） vs（5.6±1.52）],the differences between the 2 groups were all of statistical significance（P0.05）. Conclusions The exosomes of bEnd. 3 cells can affect the cell viability of neural stem cells.