细胞外泌体的分离提取标准化及临床转化进展

Advances in isolation and extraction standardization and clinical translation of exosomes

外泌体是一类由活细胞分泌的具有磷脂双分子层结构的囊泡,其内携带RNA、DNA、蛋白质、脂质等多种信号分子,起到细胞间信号传递的作用,从而调控生理病理过程。外泌体的提取技术包括差速离心技术、等密度梯度离心技术、尺寸排阻色谱技术、超滤技术、聚合物共沉淀技术、免疫亲和技术、微流控分离技术等,这些提取技术各有优缺点,目前国际上尚未统一外泌体的提取标准;而且不同细胞来源及培养条件下的外泌体其特异性蛋白和遗传物质表达有所不同,从而表现特征和功能也存在差异;因此,目前外泌体多用于临床前研究,广泛应用于临床尚存在困难和挑战。本文总结了外泌体在现阶段研究领域中的进展,归纳了不同细胞来源外泌体的特征和应用、优缺点以及面临的挑战,以期帮助研究者更好地了解和掌握外泌体的性能;同时建议需要完善外泌体提取和制备的标准化流程为外泌体的临床转化和应用提供参考。

Exosomes are phospholipid bilayer vesicles secreted by living cells that can carry a variety of signaling molecules,such as RNA,DNA,protein,and lipids.Exosomes play a role in the transmission of signaling molecules between cells,thus regulating many physiological and pathological processes.The methods of extracting exosomes include differential centrifugation,density gradient centrifugation,exclusion chromatography,ultrafiltration,coprecipitation,polymer immune affinity,microfluidic separation technology,etc.Each of these extraction technologies has advantages and disadvantages;however,there is no unified international standard.In addition,the expression of specific proteins and genetic material of exosomes from different cell sources are different;thus,their expression characteristics and functions are also distinctive.Based on this situation,research on exosomes is limited to preclinical studies,and difficulties and challenges still exist in clinical application.This paper summarizes the progress of research in the field of exosomes,to understand the characteristics,modification and application of exosomes from different cell sources,and to summarize their advantages and disadvantages as well as challenges,which can help researchers better understand and master the performance of exosomes.Furthermore,improvement of standard procedures in the extraction and manufacturing of exosomes is important,as it will provide a reference for researchers to carry out exosome-related translational clinical research.