# The new composition of circulating microvesicles: optimized protocols and reassessment of their characteristics and physiological functions

**Authors:** Chen Zhang, Jiajia Hu, Yifan Shi, Yang Feng, Zeyang Li, Zi Dong, Yiding Tang, Guang Ning, Zhengting Wang, Guorui Huang

PMC · DOI: 10.1093/lifemedi/lnaf017 · 2025-06-11

## TL;DR

This paper shows that centrifugation speed affects microvesicle composition, with most being platelet-derived at lower speeds, impacting their clinical and functional interpretations.

## Contribution

The study identifies 3000 g as a critical centrifugation speed for isolating platelet-derived microvesicles, clarifying their origin and function.

## Key findings

- Centrifugation at 3000 g yields MVs where over 80% are platelet-derived.
- Higher centrifugation speeds result in MVs with only about 20% platelet origin.
- Platelet-derived MVs are linked to procoagulation activity and clinical diagnosis.

## Abstract

Microvesicles (MVs) have convenient clinical applications and play functional roles in cellular signal transduction. Although the clinical importance of MVs is being increasingly recognized, the current diversity of isolated protocols results in a heterogeneous population of their unknown origins, even expands to uncertain functions. Here, we systematically investigated the composition of MVs at different centrifugal speed intervals and discovered that centrifugation at 3000 g is critical in determining the composition of MVs. We observed that platelet-derived particles accounted for more than 80% of MVs under 3000 g, while only about 20% of MVs were obtained over 3000 g. The discovery that more than 80% of platelet-derived MVs sheds new light on their function, including procoagulation activity and clinical diagnosis, etc. Our work not only optimizes the method for MVs isolation but also clarifies the physiological functions and characteristics that should be attributed to platelets rather than MVs. Consequently, these findings will derive new conceptualizations regarding MVs’ composition and function.

## Full-text entities

- **Genes:** NLRP3 (NLR family pyrin domain containing 3) [NCBI Gene 114548] {aka AGTAVPRL, AII, AVP, C1orf7, CIAS1, CLR1.1}, Actb (actin, beta) [NCBI Gene 11461] {aka Actx, E430023M04Rik, beta-actin}, Tfrc (transferrin receptor) [NCBI Gene 22042] {aka 2610028K12Rik, CD71, E430033M20Rik, Mtvr1, TFR, TFR1}, IL1B (interleukin 1 beta) [NCBI Gene 3553] {aka IL-1, IL1-BETA, IL1F2, IL1beta}, Nlrp3 (NLR family, pyrin domain containing 3) [NCBI Gene 216799] {aka AGTAVPRL, AII/AVP, Cias1, FCAS, FCU, MWS}, PECAM1 (platelet and endothelial cell adhesion molecule 1) [NCBI Gene 5175] {aka CD31, CD31/EndoCAM, GPIIA', PECA1, PECAM-1, endoCAM}, Il1b (interleukin 1 beta) [NCBI Gene 16176] {aka IL-1beta, Il-1b}, Gypa (glycophorin A) [NCBI Gene 14934] {aka CD235a, GPA}, CDH5 (cadherin 5) [NCBI Gene 1003] {aka 7B4, CD144}, MAEA (macrophage erythroblast attacher, E3 ubiquitin ligase) [NCBI Gene 10296] {aka EMLP, EMP, GID9, HLC-10, P44EMLP, PIG5}, Cox4i1 (cytochrome c oxidase subunit 4I1) [NCBI Gene 12857] {aka COX, COX IV-1, COXIV, Cox4, Cox4a, IV-1}
- **Diseases:** hepatocellular carcinoma (MESH:D006528), vascular injury (MESH:D057772), lung (MESH:D008171), thrombus (MESH:D013927), blood coagulation (MESH:D001778), cardiovascular disease (MESH:D002318), CAD (MESH:D003324), inflammation (MESH:D007249), tumor metastasis (MESH:D009362), cancer (MESH:D009369), ALI (MESH:D055371), sickle (MESH:D000755), gastric adenocarcinoma (MESH:D013274)
- **Chemicals:** copper (MESH:D003300), penicillin (MESH:D010406), MTT (MESH:C070243), streptomycin (MESH:D013307), Hematoxylin (MESH:D006416), LPS (MESH:D008070), Eosin (MESH:D004801), SDS (MESH:D012967), lipids (MESH:D008055), H&amp;E (MESH:D006371), carbon (MESH:D002244), PVDF (MESH:C024865), phosphotungstic acid (MESH:D010772), DMEM (-), CO2 (MESH:D002245), paraformaldehyde (MESH:C003043)
- **Species:** Homo sapiens (human, species) [taxon 9606], Escherichia coli O111 (serogroup) [taxon 1055535], Mus musculus (house mouse, species) [taxon 10090]
- **Cell lines:** C57BL/6 — Mus musculus (Mouse), Transformed cell line (CVCL_C0MU), AGS — Homo sapiens (Human), Gastric adenocarcinoma, Cancer cell line (CVCL_0139), SK-Hep1 — Homo sapiens (Human), Liver and intrahepatic bile duct epithelial neoplasm, Cancer cell line (CVCL_0525), HUVEC — Homo sapiens (Human), Finite cell line (CVCL_2959), Hela — Homo sapiens (Human), Human papillomavirus-related endocervical adenocarcinoma, Cancer cell line (CVCL_0030), iBMDM — Mus musculus (Mouse), Transformed cell line (CVCL_C0MS)

## Figures

7 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12277571/full.md

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Source: https://tomesphere.com/paper/PMC12277571