A protocol for isolating and imaging large extracellular vesicles or midbody remnants from mammalian cell culture

被引：1

作者：

Park, Sungjin ^{[1
]}

Patel, Smit A. ^{[1
]}

Torr, Elizabeth E. ^{[1
]}

Dureke, Ashley-Grace N. ^{[1
]}

Mcintyre, Alina M. ^{[1
]}

Skop, Ahna R. ^{[1
]}

机构：

[1] Univ Wisconsin, Dept Genet, Madison, WI 53706 USA

来源：

STAR PROTOCOLS | 2023年 / 4卷 / 04期

基金：

美国国家卫生研究院;

关键词：

CYTOKINESIS;

D O I：

10.1016/j.xpro.2023.102562

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Traditionally, midbody remnants (MBRs) are isolated from cell culture medium using ultracentrifugation, which is expensive and time consuming. Here, we pre-sent a protocol for isolating MBRs or large extracellular vesicles (EVs) from mammalian cell culture using either 1.5% polyethylene glycol 6000 (PEG6000) or PEG5000-coated gold nanoparticles. We describe steps for growing cells, col-lecting media, and precipitating MBRs and EVs from cell culture medium. We then detail characterization of MBRs through immunofluorescent antibody stain-ing and immunofluorescent imaging.

引用

页数：15

共 50 条

[21] Reporter mice for isolating and auditing cell type-specific extracellular vesicles in vivo
McCann, James, V
Bischoff, Steven R.
Zhang, Yu
Cowley, Dale O.
Sanchez-Gonzalez, Veronica
Daaboul, George D.
Dudley, Andrew C.
GENESIS, 2020, 58 (07)
[22] A class of large cell-like extracellular vesicles
D'Souza-Schorey, Crislyn
Di Vizio, Dolores
NATURE CELL BIOLOGY, 2025, 27 (03) : 372 - 374
[23] Large-scale mammalian cell culture
Hu, WS
Aunins, JG
CURRENT OPINION IN BIOTECHNOLOGY, 1997, 8 (02) : 148 - 153
[24] 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):
[25] Diversity of extracellular vesicles derived from calli, cell culture and apoplastic fluid of tobacco
Kocholata, Michaela
Maly, Jan
Krizenecka, Sylvie
Janouskova, Olga
SCIENTIFIC REPORTS, 2024, 14 (01):
[26] A novel approach for enumeration of extracellular vesicles from crude and purified cell culture samples
Kruse, Thomas
Schneider, Samuel
Reger, Lucas Nik
Kampmann, Markus
Reif, Oscar-Werner
ENGINEERING IN LIFE SCIENCES, 2022, 22 (3-4): : 334 - 343
[27] Size Exclusion Chromatography for Separating Extracellular Vesicles from Conditioned Cell Culture Media
Jones, Madison T.
Manioci, Samantha W.
Russell, Ashley E.
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, 2022, (183):
[28] Metabolic signature of extracellular vesicles depends on the cell culture conditions
Palviainen, Mari
Saari, Heikki
Karkkainen, Olli
Pekkinen, Jenna
Auriola, Seppo
Yliperttula, Marjo
Puhka, Maija
Hanhineva, Kati
Siljander, Pia R-M
JOURNAL OF EXTRACELLULAR VESICLES, 2019, 8 (01):
[29] Isolating high-purity extracellular vesicles from human biofluids for proteome analysis
Ter-Ovanesyan, Dmitry
Trieu, Wendy
Iyer, Siddharth
Gilboa, Tal
Nickitina, Adele
Shami-Shah, Adnan
Church, George M.
Walt, David R.
Ahmad, Rushdy
Budnik, Bogdan
MOLECULAR & CELLULAR PROTEOMICS, 2022, 21 (08) : S63 - S63
[30] A standarized protocol for evaluation of large extracellular vesicles using the attuneTM NXT system
Lugo-Gavidia, Leslie Marisol
Burger, Dylan
Bosio, Erika
Robinson, Sandi
Nolde, Janis
Carnagarin, Revathy
Matthews, Vance B.
Schlaich, Markus P.
JOURNAL OF IMMUNOLOGICAL METHODS, 2021, 499

← 1 2 3 4 5 →