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

Traditionally, midbody remnants (MBRs) are isolated from cell culture medium using ultracentrifugation, which is expensive and time consuming. Here, we present a protocol for isolating MBRs or large extracellular vesicles (EVs) from mammalian cell culture using either 1.5% polyethylene glycol 6000 (...

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Detalles Bibliográficos
Autores principales: Park, Sungjin, Patel, Smit A., Torr, Elizabeth E., Dureke, Ashley-Grace N., McIntyre, Alina M., Skop, Ahna R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10500451/
https://www.ncbi.nlm.nih.gov/pubmed/37690025
http://dx.doi.org/10.1016/j.xpro.2023.102562
Descripción
Sumario:Traditionally, midbody remnants (MBRs) are isolated from cell culture medium using ultracentrifugation, which is expensive and time consuming. Here, we present 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, collecting media, and precipitating MBRs and EVs from cell culture medium. We then detail characterization of MBRs through immunofluorescent antibody staining and immunofluorescent imaging.

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