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
收藏
页数：15
相关论文
共 50 条
  • [31] LARGE EXTRACELLULAR VESICLES FROM PREECLAMPSIA DISRUPT THE INTEGRITY OF BRAIN ENDOTHELIAL CELL MONOLAYERS
    Coxca, Jose Martin Murrieta
    Henao-Restrepo, Julian A.
    Markert, Udo R.
    Morales-Prieto, Diana M.
    PLACENTA, 2024, 154 : E56 - E56
  • [32] A standardised protocol for the evaluation of small extracellular vesicles in plasma by imaging flow cytometry
    Armitage, J. D.
    Tan, D. B. A.
    Cha, L.
    Clark, M.
    Gray, E. S.
    Fuller, K. A.
    Moodley, Y. P.
    JOURNAL OF IMMUNOLOGICAL METHODS, 2019, 468 : 61 - 66
  • [33] Monitoring of extracellular TCA cycle intermediates in mammalian cell culture
    Ritter, Joachim B.
    Genzel, Yvonne
    Reichl, Udo
    CELL TECHNOLOGY FOR CELL PRODUCTS, 2007, : 603 - +
  • [34] Modeling the coupled extracellular and intracellular environments in mammalian cell culture
    Meshram, Mukesh
    Naderi, Saeideh
    McConkey, Brendan
    Ingalls, Brian
    Scharer, Jeno
    Budman, Hector
    METABOLIC ENGINEERING, 2013, 19 : 57 - 68
  • [35] Isolation and Characterization of Extracellular Vesicles from Arabidopsis thaliana Cell Culture and Investigation of the Specificities of Their Biogenesis
    Yugay, Yulia
    Tsydeneshieva, Zhargalma
    Rusapetova, Tatiana
    Grischenko, Olga
    Mironova, Anastasia
    Bulgakov, Dmitry
    Silant'ev, Vladimir
    Tchernoded, Galina
    Bulgakov, Victor
    Shkryl, Yury
    PLANTS-BASEL, 2023, 12 (20):
  • [36] Optimizing of a suitable protocol for isolating tissue-derived extracellular vesicles and profiling small RNA patterns in hepatocellular carcinoma
    Yang, Wenjing
    Liu, Yu
    Wang, Jiyan
    Liu, Te
    Tian, Tongtong
    Li, Tong
    Ding, Lin
    Chen, Wei
    Wang, Hao
    Zhu, Jie
    Zhang, Chunyan
    Pan, Baishen
    Zhou, Jian
    Fan, Jia
    Wang, Beili
    Yang, XinRong
    Guo, Wei
    LIVER INTERNATIONAL, 2024, 44 (10) : 2672 - 2686
  • [37] Equipment for Large-Scale Mammalian Cell Culture
    Ozturk, Sadettin S.
    MAMMALIAN CELL CULTURES FOR BIOLOGICS MANUFACTURING, 2014, 139 : 69 - 92
  • [38] Shedding of extracellular membrane vesicles from brain cells in culture
    Schiera, Gabriella
    Proia, Patrizia
    Lo Cicero, Alessandra
    Saladino, Patrizia
    Savettieri, Giovanni
    Di Liegro, Italia
    INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE, 2009, 24 : S53 - S53
  • [39] Comparison of methods for isolating extracellular vesicles from human breast milk for analyzing miRNA variation
    Myers, K. R.
    DeHoff, P.
    Laurent, L. C.
    Non, A. L.
    AMERICAN JOURNAL OF HUMAN BIOLOGY, 2021, 33
  • [40] Comparative evaluation of methods for isolating small extracellular vesicles derived from pancreatic cancer cells
    Wang, Jie-Min
    Li, Yong-Jiang
    Wu, Jun-Yong
    Cai, Jia-Xin
    Wen, Jing
    Xiang, Da-Xiong
    Hu, Xiong-Bin
    Li, Wen-Qun
    CELL AND BIOSCIENCE, 2021, 11 (01):
12345