不同复苏方式对冻存外泌体的影响

doi:10.3969/j.issn.1006-7795.2023.04.021

首都医科大学学报 ›› 2023, Vol. 44 ›› Issue (4): 639-644.doi: 10.3969/j.issn.1006-7795.2023.04.021

不同复苏方式对冻存外泌体的影响

肖俐², 罗淑敏¹, 徐芳¹, 陈静¹, 邢恩鸿^2*#, 李伟华^1*#

1.首都医科大学附属北京佑安医院北京肝病研究所，北京 100069；2.承德医学院附属医院检验科，河北承德 067000

收稿日期:2022-12-26 出版日期:2023-08-21 发布日期:2023-07-26
通讯作者: 邢恩鸿, 李伟华 E-mail:xeh001@sina.com；liweihua@ccmu.edu.cn
基金资助:
国家自然科学基金资助项目（82274447），北京市自然科学基金资助项目（7212172），北京市高层次公共卫生技术人才建设资助项目（2022-2-024），北京市属医学科研院所公益发展改革试点项目（2019-6，2021-10）

Effects of different resuscitation methods on frozen exosomes

Xiao Li²，Luo Shumin¹，Xu Fang¹，Chen Jing¹，Xing Enhong^2*#，Li weihua^1*#

1.Institute of liver Diseases，Beijing Youan Hospital，Capital Medical University，Beijing 100069，China；2.Clinical Laboratory，Affiliated Hospital of Chengde Medical College，Chengde 067000，Hebei Province，China

Received:2022-12-26 Online:2023-08-21 Published:2023-07-26
Supported by:
This study was supported by National Natural Science Foundation of China（82274447），Natural Science Foundation of Beijing (7212172)，Special Project for the Construction of High-Level Health Technical Personnel in Beijing Health System (2022-2-024)，Public Welfare Development and Reform Pilot Project of Beijing Municipal Medical Research Institutes(2019-6，2021-10).

摘要/Abstract

摘要： 目的比较4 ℃、37 ℃和室温三种不同复苏方式对冻存外泌体的影响，明确冻存外泌体的最佳复苏方式。方法将冻存血浆分成三组：4 ℃冰箱过夜融化复苏组、37 ℃水浴箱融化复苏组、室温融化复苏组。采用超速离心法提取外泌体；使用透射电镜检测外泌体大小、形态结构；Amnis量化成像流式细胞仪检测外泌体的数量和所占总粒子百分比（纯度）。结果三种复苏方式中，37 ℃水浴箱融化复苏组得到外泌体的数量和百分比最高，与4 ℃冰箱融化复苏组和室温融化复苏组比较，组间差异均有统计学意义（P＜0.01）。4 ℃冰箱过夜融化复苏组获得外泌体的数量和百分比最低。同时发现，在人的血浆外泌体中，CD9阳性外泌体的数量明显高于CD63阳性外泌体和CD81阳性外泌体，差异具有统计学意义（P＜0.05）。结论 37 ℃水浴箱融化复苏获得外泌体的数量和百分比最高，是最佳的外泌体复苏方式。在人血浆外泌体中，CD9阳性外泌体的数量及百分比最高。

关键词: 外泌体, 复苏, 活性

Abstract:

Objective To compare the effects of three resuscitation methods (4 ℃, 37 ℃ and room temperature) on frozen exosomes，to clarify the optimal resuscitation way for frozen exosomes. Methods Frozen plasma was thawed as below three methods: 4 ℃ refrigerator overnight, 37 ℃water bath box, and room temperature. Exosomes were isolated by ultracentrifugation, transmission electron microscope observed exsomes size and morphology, Amnis imaging flow cytometry detected exosome quantity and percentage (purity). Results Among the three resuscitation methods, 37 ℃ water bath resuscitation group obtained the highest number and percentage of exosomes, which were statistically higher compared to the 4 ℃ refrigerator resuscitation group and the room temperature resuscitation group (P<0.01). The number and percentage of exosomes obtained from the 4 ℃ refrigerator overnight resuscitation group were the lowest. At the same time, it was found that the number of CD9 positive exsomes in human plasma was significantly higher than that of CD63 positive and CD81 positive exsomes, with statistically significant difference (P<0.05).Conclusions The 37 ℃ water bath resuscitation method was the best method for frozen exosomes. In human plasma exosomes, CD9 positive exosomes was the highest quantity and percentage subgroup.

Key words: exosomes, resuscitation, activity

中图分类号:

R331

肖俐, 罗淑敏, 徐芳, 陈静, 邢恩鸿, 李伟华. 不同复苏方式对冻存外泌体的影响[J]. 首都医科大学学报, 2023, 44(4): 639-644.

Xiao Li, Luo Shumin, Xu Fang, Chen Jing, Xing Enhong , Li weihua. Effects of different resuscitation methods on frozen exosomes[J]. Journal of Capital Medical University, 2023, 44(4): 639-644.

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不同复苏方式对冻存外泌体的影响

Effects of different resuscitation methods on frozen exosomes

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