离心过滤法提取外切体的实验研究

目的:利用外切体的物理特性,用基于孔径分离的生物膜离心过滤法制备外切体,以降低其研究及应用的技术门槛. 方法:以树突状细胞的培养上清作为外切体的来源,分别用传统的超高速密度梯度离心法和生物膜离心过滤法制备外切体,再利用透射电镜进行两者之间形态学鉴定,聚丙烯酰胺凝胶电泳进行蛋白质成分的鉴定. 结果:两种方法的提取物在超微结构及形态方面非常相像,其蛋白质谱几乎一致. 结论:基于生物膜分离技术的离心过滤法可制备出外切体.

Isolation of exosomes by bio-membrane centrifugal filtering

AIM: To isolate exosomes by their physical diameters using bio membrane centrifugal filtering so as to lower the research and application technical threshold of exosomes. METHODS: The exosomes from the dendritic cells (DC) culture supernatant were purified and isolated by both differential centrifugation and centrifugal filtering, respectively. Transsimion electron microscopy (TEM) was used to identify the morphology of the two products and sodium dodecyl sulphate PolyAcrylamide Gel Electrophoresis (SDS PAGE) was employed to analyze their proteins. RESULTS: The 2 products have similar morphology and ultrastructure, and almost the same protein profile. CONCLUSION: Exosomes can be obtained by the centrifugal filtering method.