RRC ID 78245
Author Ito F, Yanatori I, Kato K, Toyokuni S.
Title Protocol for the isolation of GFP-expressing ferroptosis-dependent extracellular vesicles in in vitro cell culture models.
Journal STAR Protoc
Abstract Extracellular vesicles (EVs) are complex structures that transport various DNA, RNA, and protein. Recently, new EV secretion mechanisms have been identified through the iron regulatory system in mammalian cells. We revealed that ferroptosis increases EV secretion, which is named ferroptosis-dependent EVs (FedEVs). Here, we describe a step-by-step procedure to isolate GFP-expressing FedEVs for in vitro analysis. The FedEVs are further analyzed by imaging and flow cytometry analysis. For complete details on the use and execution of this protocol, please refer to Ito et al.1.
Volume 5(1)
Pages 102892
Published 2024-2-14
DOI 10.1016/j.xpro.2024.102892
PII S2666-1667(24)00057-1
PMID 38363686
PMC PMC10877193
MeSH Animals Cell Culture Techniques Extracellular Vesicles* / chemistry Ferroptosis* / genetics Mammals Proteins / metabolism
Human and Animal Cells 293T(RCB2202) THP-1(RCB1189)
DNA material pCMV-VSV-G-RSV-Rev (RDB04393) pCAG-HIVgp (RDB04394)