人脐带间充质干细胞来源外泌体的提取与鉴定

Extraction and identification of exosomes from human umbilical cord mesenchymal stem cells

目的:探讨人脐带间充质干细胞(HUCMSC)来源外泌体的有效提取方法。方法:通过超速离心法从脐带干细胞培养上清液中分离外泌体,采用蛋白免疫印迹法、透射电镜对提取到的外泌体进行鉴定。结果:分离获得的外泌体特异富集蛋白CD9、TSG101等,透射电镜下可见其呈杯盘状结构,直径约30~100 nm,有完整的膜结构,内含低密度物质。结论:HUCMSC分泌的外泌体,具有外泌体的经典结构及典型特征,并且易提取,操作方便,为未来脐带干细胞外泌体作为载体进行药物递送提供了实验依据。

Objective:To explore the effective extraction method of exosomes from human umbilical cord mesenchymal stem cells.Methods:Exosomes were isolated from the culture supernatant of umbilical cord stem cells by ultracentrifugation,the exosomes were identified by Western blot and transmission electron microscopy.Results:The exosome specific enriched proteins CD9 and TSG101 were isolated,under transmission electron microscope,it is a cup-shaped structure with a diameter of about 30~100 nm,it has a complete membrane structure and contains low-density substances.Conclusion:The exosomes secreted by HUCMSC have the classical structure and typical characteristics of exosomes,and are easy to extract and operate.This study provides an experimental basis for the future drug delivery by umbilical cord stem cell exosomes.