外泌体之家 | 细胞外膜泡领域核心平台—exosomes & microvesicles—小膜泡大作用
标题: 干细胞的外泌体综述 [打印本页] 作者: zhuyiping 时间: 2015-7-29 16:38 标题: 干细胞的外泌体综述 有关间充质干细胞外泌体的综述 作者: zhuyiping 时间: 2015-8-5 16:10
1.For the isolation of exosomes from human umbilical cord Wharton’s jelly-MSCs and human dermal fibroblasts, serum-free culture medium conditioned for 24 hours was filtered (0.2 m) and concentrated by ultrafiltration device with 100-kDa cutoff (Millipore).
2.Exosomes in CM were precipitated with one-third volume of polyethylene glycol buffer (33.4% PEG 4000, 50 mmol/L HEPES [pH 7.4], 1 mol/L NaCl) overnight at 4°C, followed by centrifugation at 12 000 g for 5 minutes and resuspension in PBS (pH 7.4).
3.Exosomes in polyethylene glycol–precipitated fraction were further purified by S200 size-exclusion chromatography. A 75-L sample was applied on a S200 column (Clontech, Mountain View, CA)pre-equilibrated with PBS by spinning at 700 g for 5 minutes, and the exosomal fraction was subsequently eluted in the flow-through by
centrifugation at 700 g for 5 minutes.
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