Effects of different resuscitation methods on frozen exosomes

doi:10.3969/j.issn.1006-7795.2023.04.021

Abstract

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

CLC Number:

R331

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.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2023.04.021

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2023/V44/I4/639

References

［1］ Tenchov R, Sasso J M, Wang X M, et al. Exosomes─nature's lipid nanoparticles, a rising star in drug delivery and diagnostics［J］. ACS Nano, 2022, 16(11): 17802-17846.
［2］ Liang Y,Duan L, Lu J,et al.Engineering exosomes for targeted drug delivery［J］.Theranostics, 2021,11(7):3183-3195.
［3］ Cully M. Exosome-based candidates move into the clinic［J］. Nat Rev Drug Discov, 2021, 20(1): 6-7.
［4］ Xu Z J, Zeng S S, Gong Z C, et al. Exosome-based immunotherapy: a promising approach for cancer treatment［J］. Mol Cancer, 2020, 19(1): 160.
［5］ Pegtel D M, Gould S J. Exosomes［J］. Annu Rev Biochem, 2019, 88: 487-514.
［6］ Cheng Y R, Zeng Q Y, Han Q, et al. Effect of pH, temperature and freezing-thawing on quantity changes and cellular uptake of exosomes［J］. Protein Cell, 2019, 10(4): 295-299.
［7］ Tertel T, Görgens A, Giebel B. Analysis of individual extracellular vesicles by imaging flow cytometry［J］. Methods Enzymol, 2020, 645: 55-78.
［8］ Zhang Y, Bi J Y, Huang J Y, et al. Exosome: a review of its classification, isolation techniques, storage, diagnostic and targeted therapy applications［J］. Int J Nanomedicine, 2020, 15: 6917-6934.
［9］李榕, 陈静, 李传云, 等. 过滤对超速离心法提取外泌体的影响［J］. 首都医科大学学报, 2021, 42(6): 1007-1013.
［10］ Kalluri R, LeBleu V S. The biology, function, and biomedical applications of exosomes［J］. Science, 2020, 367(6478): eaau6977.
［11］阚宠, 黎建昌, 黄小程, 等. 外泌体相关跨膜蛋白上调整合素促进肝细胞癌中细胞迁移的研究［J］. 右江医学, 2020, 48(3): 188-193.
［12］ Böker K O, Lemus-Diaz N, Rinaldi Ferreira R, et al. The impact of the CD9 tetraspanin on lentivirus infectivity and exosome secretion［J］. Mol Ther, 2018, 26(2): 634-647.
［13］ Khan N Z, Cao T X, He J Y, et al. Spinal cord injury alters microRNA and CD81+ exosome levels in plasma extracellular nanoparticles with neuroinflammatory potential［J］. Brain Behav Immun, 2021, 92: 165-183.
［14］ Yanatori I, Richardson D R, Dhekne H S, et al. CD63 is regulated by iron via the IRE-IRP system and is important for ferritin secretion by extracellular vesicles［J］. Blood, 2021, 138(16): 1490-1503.

[1]	Yang Sen, Wang Ling, Liu Peng, Jin Chaoling, Li Honglei, Fu Liping. Comparison of continuous-bed-motion and step-and-shoot acquisition modes on ¹⁸F-FDG PET/CT imaging [J]. Journal of Capital Medical University, 2022, 43(6): 854-860.
[2]	Lu Yan, Li Zhen, Wu Yue, Zhang Jian, Jiang Huizhong, Wang Ningli. Study of the correlation between biomarkers of Alzheimer's disease in aqueous humor and age in normal people [J]. Journal of Capital Medical University, 2017, 38(3): 461-464.
[3]	Shi Luping;Xu Meng;Liu Jie;Wang Jun. Correlation of Volume-sensitivity with Potassium Channels in Ventricular Myocyte of Rats [J]. Journal of Capital Medical University, 2008, 29(5): 585-588.
[4]	Xie Miaorong;Shen Luhua. The Impact of Nonmyocytes on the Expression of α-and β-Myosin Heavy Chain mRNA of Myocytes [J]. Journal of Capital Medical University, 2007, 28(4): 427-429,433.
[5]	Xie Miaorong;Shen Luhua. The Impact of Nonmyocytes Conditioned Media on the Biological Process of Apoptosis in Myocytes [J]. Journal of Capital Medical University, 2007, 28(4): 430-433.
[6]	Wang Wei;Huang Haixia;Zhang Lijuan;Ma Tao;Liu Ping;Wei Hua;Niu Weizhen. Mechanosensitivity of ATP-Sensitive Potassium Channels in Rat Colonic Smooth Muscle Cells [J]. Journal of Capital Medical University, 2007, 28(3): 341-344.
[7]	Gao Ying;Yang Xinchun;Zhou Peng;Liu Xiulan. Study of Electrical Heterogeneity of L-type Calcium Current in the Canine Ventricular Myocytes [J]. Journal of Capital Medical University, 2007, 28(1): 81-84.
[8]	Xing Yan;Jiao Xiping;Wang Baoguo . Correlation Between Partial CO₂ Rebreathing Technique and Transesophageal Doppler for Cardiac Output Monitoring During Controlled Hypotension [J]. Journal of Capital Medical University, 2005, 26(2): 216-218.
[9]	Zhang Lianguo;Li Xiaojing;Liu Mingfang;Zhang Lin;Wang Shuo;Xiao Han;Nie Xiuhong. Characteristics of Peripheral Lymphocyte in Patients with Severe Acute Respiratory Syndrome [J]. Journal of Capital Medical University, 2003, 24(4): 389-391.
[10]	Zhang Lianguo;Liu Mingfang;Nie Xiuhong;Zhang Lin;Wang Shuo;Li Xiaojing;Xiao Han;Yang Qiang. Effects of Platelete Counts on Prognosis in Patients with Severe Acute Respiratory Syndrome [J]. Journal of Capital Medical University, 2003, 24(4): 453-456.
[11]	Su Li;Tian Ding;Wan Suigui;Liu Congyan;Xu Juan . Study on Cord Blood CD34⁺ Cell in Short-term Culture in Vitro Amplification [J]. Journal of Capital Medical University, 2002, 23(3): 218-221.
[12]	Yan Yan;Liu Xuezong;Wang Xianming;Niu Weizhen. Comparison of Numerical Methods to Process Data of Myocytes Ca²⁺ Fluointersity [J]. Journal of Capital Medical University, 2002, 23(1): 20-23.
[13]	Ma Yaluan;Gao Tianxiang;Wang Jianing;Qi Yahui;Yin Hong;Feng Guifen . Antisense c-myc Oligodeoxynucleotides Inhibited Proliferation, Migration of Vascular Smooth Muscle Cells and Induced Apoptosis by Downregulation of c-myc Expression [J]. Journal of Capital Medical University, 1998, 19(3): 205-208.
[14]	Zhang Wanjiang;Zhao Zhiwei;Wang Rong;Sheng Shuli. Therapeutic Effects of Calcitonin Gene Related Peptide on Arrhythmias Caused by Ouabain and Aconitine in Rats [J]. Journal of Capital Medical University, 1994, 15(3): 204-208.
[15]	Jiao Xiaohui;Chen Xiaoliang;Han Daying. Protective Action of the Vescular Endothelial Cells on Tension of the Smooth Muscle [J]. Journal of Capital Medical University, 1994, 15(2): 100-102.