中国组织工程研究2018，Vol.22Issue(9)：1382-1388,7.DOI:10.3969/j.issn.2095-4344.0468
人脐带间充质干细胞来源外泌体提取方法的比较
Comparison of exosome extracting methods from human umbilical cord mesenchymal stem cells
摘要
Abstract
BACKGROUND: Cell-free stem cell therapy has been an issue of concern, but there is no conclusion on how to extract high-quality exosomes. OBJECTIVE: To extract exosomes from human umbilical cord mesenchymal stem cells by using three different methods, and then to screen the optimal method. METHODS: Exosomes were extracted from human umbilical cord mesenchymal stem cells by using the Total Exosome Isolation test kit, Exo Quick test kit and differential ultracentrifugation method, respectively. Then, transmission electron microscopy was used for morphological observations, BCA was utilized to quantify the protein, and western blot assay was applied to detect surface markers CD9, CD81 and CD63. RESULTS AND CONCLUSION: Extraction of exosomes was completed by all the three methods, and round or oval membranous vesicles were observed under the transmission electron microscope. The protein content and purity of exosomes was highest in the differential ultracentrifugation group, followed by the Exobiology Quick kit group, and lowest in the Total Exosome Isolation kit group, and there were significant differences among the three groups (P < 0.05). Under the same protein concentration, surface specific markers, CD81, CD63 and CD9, were expressed highest in the differential ultracentrifugation group, followed by the Exobiology Quick kit group, and lowest in the Total Exosome Isolation kit group. The operating time was significantly lower in the Exobiology Quick kit group compared with the other two groups (P < 0.05). To conclude, despite a longer operating time, the differential ultracentrifugation method is a rational method to extract enough exosomes with relative high purity.关键词
外泌体/提取方法/培养基/干细胞/Exo Quic-TC试剂盒法/差速超速离心法/Total Exosome Isolation试剂盒法/人脐带间充质干细胞/干细胞治疗/"无细胞"替代疗法/国家自然科学基金分类
医药卫生引用本文复制引用
郭莹,牛勃,王秀伟,牛玉虎,王莉,周楠,李佰一,王振东,张蘋,高亚杰..人脐带间充质干细胞来源外泌体提取方法的比较[J].中国组织工程研究,2018,22(9):1382-1388,7.基金项目
国家自然科学基金(81370312) （81370312）
海南省自然科学基金(812180) （812180）
三亚市医疗卫生科技创新项目(2014 YW01) （2014 YW01）
the National Natural Science Foundation of China,No.81370312 （）
the Natural Science Foundation of Hainan Province,No.812180 （）
the Scientific Innovation Project for Medical Treatment and Public Health in Sanya City,No.2014 YW01 （）
