淋巴瘤来源外泌体的提取方法比较与鉴定

Comparison of isolation methods of lymphoma-derived exosomes

目的比较淋巴瘤来源外泌体的3种提取方法[聚乙二醇(PEG)沉淀法、exoEasy试剂盒、超速离心法],并鉴定其形态及生物学特征。方法采用PEG沉淀法、exoEasy试剂盒、超速离心法提取外泌体,在透射电子显微镜下观察外泌体形态,利用Western blot法检测3种方法提取的外泌体蛋白表达特点。结果PEG沉淀法、超速离心法提取的外泌体能表达特征蛋白CD63、CD81,具备外泌体的基本形态特征,其中超速离心法提取的外泌体特征更清晰;而exoEasy试剂盒提取的外泌体特征不明显。结论PEG沉淀法适合对外泌体产量需求大的实验,超速离心法适合外泌体的功能实验,exoEasy试剂盒适合外泌体前期预实验的探索使用。可根据不同研究目的选择不同的提取方法。

Objective To compare three different methods for isolation of lymphoma-derived exosomes(EXOs).Methods EXOs were extracted from lymphoma cell lines by PEG precipitation method,hypervelocity centrifugation method or exoEasy commercial kit,respectively.The morphology and protein expression of EXOs were detected by electron microscopy and Western blot,respectively.Results EXOs extracted by PEG precipitation and hypervelocity centrifugation could express the characteristic proteins CD63 and CD81,which had the classical morphological characteristics of EXOs,but the EXOs extracted by hypervelocity centrifugation were more clearly.EXOs extracted by the exoEasy kit had no significantly clear characteristics.Conclusion PEG precipitation method is suitable for EXOs isolation with large production,while hypervelocity centrifugation method is suitable for EXOs functional experiments,and the exoEasy kit is suitable for EXOs pre-experimental exploration.Different extraction methods can be used for different requirements of research.