外泌体之家 | 细胞外膜泡领域核心平台—exosomes & microvesicles—小膜泡大作用

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exosomes-free FBS的获取方法

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发表于 2015-5-8 09:32:54 | 只看该作者 回帖奖励 |倒序浏览 |阅读模式
      Standard growth medium for most cells in culture require fetal bovine serum (FBS) as a growth supplement to DMEM/RPMI. FBS is derived from bovine (cow) serum and contains a high abundance of cow-derived exosome vesicles. These exosomes can interfere or cause significant protein background issues when studying exosomes secreted from cells of interest in standard culture conditions. For exosome production, we typically modify the growth medium either serum-free media (SFM) or a combination of SFM and Insulin Transferrin-Selenium (ITS, 0.2–0.6 % (w/v)) (i.e., culture medium B) to ensure cells remain viable during exosome production. Any changes to cell growth medium supplements should be evaluated over the time-course of exosome-production (typically 24–48 h). Further methods to modify the cell growth medium (although still the possibility of bovine secreted proteins) include, 1 % (w/v) bovine serum albumin (BSA) added instead of FBS, and depletion of FCS-containing exosomes based on
ultracentrifugation (100,000 × g, 18 h), or filtration/immunodepletion (CD63 positive, exosome-sized vesicles removed, no measureable bovine microRNAs). This exosome-depleted FBS growth supplement (Exo-FBS) is commercially available (EXO-FBS-250A-1, System Biosciences).


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发表于 2015-5-11 18:43:20 | 只看该作者
很好,感谢分享
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