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  3. Protocol for the isolation of GFP-expressing ferroptosis-dependent extracellular vesicles in in vitro cell culture models

Protocol for the isolation of GFP-expressing ferroptosis-dependent extracellular vesicles in in vitro cell culture models

  • STAR Protoc. 2024 Feb 14;5(1):102892. doi: 10.1016/j.xpro.2024.102892.
Fumiya Ito 1,  Izumi Yanatori 2,  Katsuhiro Kato 3,  Shinya Toyokuni 4
Affiliations

Affiliations

  • 1 Department of Pathology and Biological Responses, Nagoya University Graduate School of Medicine, 65 Tsurumai-Cho, Showa-ku, Nagoya 466-8550, Japan; Department of Cell Biology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. Electronic address: fito1@jhu.edu.
  • 2 Department of Molecular and Cellular Physiology, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
  • 3 Department of Cardiology, Nagoya University Graduate School of Medicine, 65, Tsurumai-Cho, Showa-ku, Nagoya 466-8550, Japan.
  • 4 Department of Pathology and Biological Responses, Nagoya University Graduate School of Medicine, 65 Tsurumai-Cho, Showa-ku, Nagoya 466-8550, Japan. Electronic address: toyokuni@med.nagoya-u.ac.jp.
PMID: 38363686 DOI: 10.1016/j.xpro.2024.102892
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.

Keywords

Cancer; Cell Biology; Cell culture; Cell isolation; Flow Cytometry.

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