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采用高速离心及ExoQuick-TC法提取小鼠骨髓间充质干细胞来源外泌体比较及鉴定
引用本文:韩金秀,李宏远,撒亚莲,严丹,贺继刚.采用高速离心及ExoQuick-TC法提取小鼠骨髓间充质干细胞来源外泌体比较及鉴定[J].安徽医科大学学报,2017,52(4).
作者姓名:韩金秀  李宏远  撒亚莲  严丹  贺继刚
作者单位:昆明理工大学附属云南省第一人民医院心脏大血管外科,昆明,650032;昆明理工大学附属云南省第一人民医院基础研究所,昆明,650032;昆明理工大学附属云南省第一人民医院重症医学科,昆明,650032
基金项目:国家自然科学基金,云南省科技厅-昆明医科大学应用基础研究联合专项,云南省教育厅科学研究基金,中国博士后科学基金,成都医学院校级课题
摘    要:比较及鉴定超高速离心法及ExoQuick-TC法提取小鼠骨髓间充质干细胞来源外泌体.采用BCA蛋白检测可见ExoQuick-TC法提取的外泌体蛋白含量高于超高速离心法提取的蛋白含量.Exosome Antibodies,Array & ELISA Kits检测可见ExoQuick-TC法提取外泌体的纯度优于超高速离心法提取的纯度.两种方法提取的外泌体电镜下形状无差异,随机选取3个电镜视野进行计数可见ExoQuick-TC法提取外泌体数量多于超高速离心法提取外泌体. ExoQuick-TC法提取的外泌体具有更高的产量及纯度.

关 键 词:外泌体  小鼠骨髓间充质干细胞  超高速离心  ExoQuick-TC法

Comparison of ultracentrifugation and ExoQuick-TC for the identificationof exosomes extracted from mouse bone marrow mesenchymal stem cells
Abstract:To compare ultracentrifugation and ExoQuick-TC in identifying the characteristics of exosomes extracted from mouse bone marrow mesenchymal stem cells.BCA protein assay revealed that the protein contents in the ExoQuick-TC-extracted exosomes were higher than those in the ultracentrifugation-extracted exosomes.Analysis using exosome antibodies, array, and ELISA kits indicated that the purity of the ExoQuick-TC-extracted exosomes was higher that of the ultracentrifugation-extracted exosomes.Electronmicroscopy demonstrated that the morphological characteristics of the ExoQuick-TC-extracted exosomes did not significantly differ from those of the ultracentrifugation-extracted exosomes.Three electron microscope visual fields were randomly selected to count the number of exosomes.Results showed that the number of the ExoQuick-TC-extracted exosomes was more than that of the ultracentrifugation-extracted exosomes.Therefore, the production and purity of ExoQuick-TC-extracted exosomes are higher than those of ultracentrifugation-extracted exosomes.
Keywords:exosome  mouse bone marrow mesenchymal stem cells  ultra-high speed centrifuge  ExoQuick-TC
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