Storage of plasma-derived exosomes: evaluation of anticoagulant use and preserving temperatures

doi:10.1080/09537104.2024.2337255

	Storage of plasma-derived exosomes: evaluation of anticoagulant use and preserving temperatures
	Yang, Caiting 9,10; Han, Jie 6,7,8; Liu, Hai 9,10; He, Yuyu 9,10; Zhang, Zhenhua 5; Liu, Xiaochun 5; Waqas, Farooq 9,10; Zhang, Lizhong 5; Duan, Huiping 3,4; He, Jing 2,5
刊名	PLATELETS
	2024-12-31
卷号	35 期号:1 页码:9
关键词	exosome anticoagulant plasma temperature duration
ISSN号	0953-7104
DOI	10.1080/09537104.2024.2337255
通讯作者	Duan, Huiping(duanhuiping461@163.com) ; He, Jing(hejing3368@163.com) ; Dong, Li(dongli@sxu.edu.cn)
英文摘要	Exosomes carry large cargo of proteins, lipids, and nucleic acids, serving as versatile biomarkers for disease diagnosis and vehicles for drug delivery. However, up to date, no well recognized standard procedures for exosome storage were available for clinical application. This study aimed to determine the optimal storage conditions and the anticoagulants for plasma-derived exosome isolation. Fresh whole blood samples were collected from healthy participants and preserved in four different anticoagulants including sodium citrate (SC1/4), sodium citrate (SC1/9), lithium heparin (LH), or Ethylenediamine tetraacetic acid (EDTA), respectively. Exosomes were extracted from the plasma by differential ultracentrifugation and stored at three different temperatures, 4 degrees C, -20 degrees C or - 80 degrees C for a duration ranging from one week to six months. All plasma samples for storage conditions comparison were pretreated with LH anticoagulant. Exosome features including morphological characteristics, pariticles size diameter, and surface protein profiles (TSG101, CD63, CD81, CD9, CALNEXIN) were assessed by transmission electron microscopy, Nanoparticle Tracking Analysis, and Western Blotting, respectively. Exosomes preserved in LH and SC1/4 group tended to remain intact microstructure with highly abundant protein biomarkers. Exosomes stored at 4 degrees C for short time were prone to be more stable compared to thos at -80 degrees C. Exosomes stored in plasma were superior in terms of ultrastructure, size diameter and surface protein expression to those stored in PBS. In conclusion, plasma-dervied exosome characteristics strictly depend on the anticoagulants and storage temperature and duration. What is the context? Effective isolation of exosomes is a prerequisite for subsequent investigation into its involvemnt in disease development as well as potentialtherapeutic applications.Anticoagulants, storage temperature and durations might change the microscopical structure, integrity and also the stability of plasma-derived exosomes. However, no internationally recognized standard of exosome storage procedure was available for clinical use.What is new? Our finding evaluated the effect of anticoagulants and storage on plasma exosome characteristics.Exosomes isolated from plasma preserved with Li-heparin and sodium citrate (1/4) showed better physical properties and surface marker protein expression.Isolated exosomes appeared more stable in a short time for 4 degrees C compared to -80 degrees C. Storage of exosomes in plasma showed better physical properties and surface marker protein expression than in PBS.What is the impact? Our findings inform the significance of standardizing procedure of exosome isolation and preservation.
资助项目	Key Research and Development program of Shanxi province[202102130501009] ; Key Research and Development program of Shanxi province[202203021221300] ; Natural Science Foundation of Shanxi Province[202203021221300] ; Shanxi Provincial Key Laboratory of Medical Molecular Cell Biology[MMCB-OP-2023- 02] ; Four Batches Innovation Project of Invigorating Medical through Science and Technology of Shanxi Province[2023XM028] ; Scientific and Technological Innovation Programs of Higher Education Institutions in Shanxi[2019L0102]
WOS关键词	EXTRACELLULAR VESICLES
WOS研究方向	Cell Biology ; Hematology
语种	英语
WOS记录号	WOS:001205226600001
资助机构	Key Research and Development program of Shanxi province ; Natural Science Foundation of Shanxi Province ; Shanxi Provincial Key Laboratory of Medical Molecular Cell Biology ; Four Batches Innovation Project of Invigorating Medical through Science and Technology of Shanxi Province ; Scientific and Technological Innovation Programs of Higher Education Institutions in Shanxi
内容类型	期刊论文
源URL	[http://dspace.imech.ac.cn/handle/311007/94984]
专题	力学研究所_国家微重力实验室
通讯作者	Duan, Huiping; He, Jing; Dong, Li
作者单位	1.Shanxi Univ, Inst Biomed Sci, 92 Wucheng Rd, Taiyuan 030006, Peoples R China 2.Shanxi Bethune Hosp, Shanxi Acad Med Sci, 99 Longcheng St, Taiyuan 030032, Peoples R China 3.Fourth Peoples Hosp Taiyuan, 231 Xikuang St, Taiyuan 030006, Peoples R China 4.Fourth Peoples Hosp Taiyuan, TB Dept, Taiyuan, Peoples R China 5.Shanxi Med Univ, Shanxi Bethune Hosp, Tongji Shanxi Hosp, Shanxi Acad Med Sci,Hosp 3, Taiyuan, Peoples R China 6.Univ Chinese Acad Sci, Sch Engn Sci, Beijing, Peoples R China 7.Chinese Acad Sci, Inst Mech, Beijing Key Lab Engn Construct & Mechanobiol, Beijing, Peoples R China 8.Chinese Acad Sci, Ctr Biomech & Bioengn, Key Lab Micrograv, Nat Micrograv Lab, Beijing, Peoples R China 9.Shanxi Univ, Inst Biomed Sci, Taiyuan, Peoples R China 10.Shanxi Univ, Minist Educ, Shanxi Prov Key Lab Med Mol Cell Biol, Key Lab Chem Biol & Mol Engn, Taiyuan, Peoples R China
推荐引用方式 GB/T 7714	Yang, Caiting,Han, Jie,Liu, Hai,et al. Storage of plasma-derived exosomes: evaluation of anticoagulant use and preserving temperatures[J]. PLATELETS,2024,35(1):9.
APA	Yang, Caiting.,Han, Jie.,Liu, Hai.,He, Yuyu.,Zhang, Zhenhua.,...&Dong, Li.(2024).Storage of plasma-derived exosomes: evaluation of anticoagulant use and preserving temperatures.PLATELETS,35(1),9.
MLA	Yang, Caiting,et al."Storage of plasma-derived exosomes: evaluation of anticoagulant use and preserving temperatures".PLATELETS 35.1(2024):9.