Protocol for the separation of extracellular vesicles by ultracentrifugation from in vitro cell culture models

被引：34
|
作者：
Chhoy, Peter [1 ]
Brown, Caitlin W. [1 ]
Amante, John J. [1 ]
Mercurio, Arthur M. [1 ]
机构：
[1] Univ Massachusetts, Dept Mol Cell & Canc Biol, Med Sch, Worcester, MA 01605 USA
来源：
STAR PROTOCOLS | 2021年 / 2卷 / 01期
关键词：
Cancer; Cell biology; Cell culture; Cell membrane;
D O I：
10.1016/j.xpro.2021.100303
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Extracellular vesicles (EVs) play key roles in transporting key molecular constitu-ents as cargo for extracellular trafficking. While several approaches have been developed to extract EVs from mammalian cells, the specific method of EV isola-tion can have a profound effect on membrane integrity and yield. Here, we describe a step-by-step procedure to separate EVs from adherent epithelial cells using differential ultracentrifugation. Separated EVs can be further analyzed by immunoblotting, mass spectrometry, and transmission electron microscopy to derive EV yield and morphology. For complete details on the use and execution of this protocol, please refer to Brown et al. (2019).
引用
收藏
页数：11
相关论文
共 50 条
  • [1] Protocol for isolating extracellular vesicles from alveolar macrophages phagocytosing MRSA in vitro cell culture models
    Bai, Songjie
    Hu, Ruomeng
    Wen, Xuehuan
    Zhang, Wenting
    Li, Bingyu
    Yang, Jie
    Yu, Qing
    Zeng, Xianchang
    Xiu, Huiqing
    Cai, Zhijian
    Zhang, Gensheng
    STAR PROTOCOLS, 2025, 6 (01):
  • [2] Protocol for separation of fungal extracellular vesicles using ultracentrifugation from solid medium cultures
    Piffer, Alicia C.
    Reis, Flavia C. G.
    Moyrand, Frederique
    Montalvao, Bruna
    Rodrigues, Marcio L.
    Janbon, Guilhem
    Rizzo, Juliana
    STAR PROTOCOLS, 2024, 5 (02):
  • [3] Protocol for the isolation of GFP-expressing ferroptosis-dependent extracellular vesicles in in vitro cell culture models
    Ito, Fumiya
    Yanatori, Izumi
    Kato, Katsuhiro
    Toyokuni, Shinya
    STAR PROTOCOLS, 2024, 5 (01):
  • [4] Protocol Protocol for isolating leukemia-derived extracellular vesicles from the spleen of preclinical models of leukemia using ultracentrifugation
    Gargiulo, Ernesto
    Morande, Pablo Elias
    Jeyakumar, Maxmilan
    Rospape, Lucie
    Paggeti, Jeome
    Moussay, Etienne
    STAR PROTOCOLS, 2024, 5 (03):
  • [5] Salivary Extracellular Vesicles Separation: Analysis of Ultracentrifugation-Based Protocols
    Itzel, Castillejos-Garcia
    Eduardo, Martinez-Martinez
    Velia, Ramirez-Amador
    Mireya, Cisneros-Villanueva
    Alfredo, Hidalgo-Miranda
    del Pilar, Ramos-Godinez Maria
    Gabriela, Anaya-Saavedra
    ORAL DISEASES, 2024,
  • [6] In vitro culture models to study bovine mammary gland extracellular vesicles
    Silvestrelli, G.
    Ulbrich, S. E.
    Saenz-de-Juano, M.
    REPRODUCTION IN DOMESTIC ANIMALS, 2020, 55 : 29 - 30
  • [7] Enrichment of calcifying extracellular vesicles using density-based ultracentrifugation protocol
    Hutcheson, Joshua D.
    Goettsch, Claudia
    Pham, Tan
    Iwashita, Masaya
    Aikawa, Masanori
    Singh, Sasha A.
    Aikawa, Elena
    JOURNAL OF EXTRACELLULAR VESICLES, 2014, 3 (01):
  • [8] A protocol for isolating and imaging large extracellular vesicles or midbody remnants from mammalian cell culture
    Park, Sungjin
    Patel, Smit A.
    Torr, Elizabeth E.
    Dureke, Ashley-Grace N.
    Mcintyre, Alina M.
    Skop, Ahna R.
    STAR PROTOCOLS, 2023, 4 (04):
  • [9] Isolation of Extracellular Vesicles Derived from Mesenchymal Stromal Cells by Ultracentrifugation
    Ramirez-Bajo, Maria Jose
    Banon-Maneus, Elisenda
    Rovira, Jordi
    Campistol, Josep M.
    Diekmann, Fritz
    BIO-PROTOCOL, 2020, 10 (24):
  • [10] Ultracentrifugal separation, characterization, and functional study of extracellular vesicles derived from serum-free cell culture
    Hu, Hooi Ting
    Nishimura, Tamako
    Suetsugu, Shiro
    STAR PROTOCOLS, 2021, 2 (03):
12345