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.
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页数：15
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