Protocol for detection of ferroptosis in cultured cells

Magdalena B Murray; Logan B Leak; Weaverly Colleen Lee; Scott J Dixon

doi:10.1016/j.xpro.2023.102457

Protocol for detection of ferroptosis in cultured cells

STAR Protoc. 2023 Sep 15;4(3):102457. doi: 10.1016/j.xpro.2023.102457. Epub 2023 Aug 8.

Authors

Magdalena B Murray¹, Logan B Leak², Weaverly Colleen Lee², Scott J Dixon³

Affiliations

¹ Department of Biology, Stanford University, 327 Campus Dr, Stanford, CA 94305, USA. Electronic address: mbmurray@stanford.edu.
² Department of Biology, Stanford University, 327 Campus Dr, Stanford, CA 94305, USA.
³ Department of Biology, Stanford University, 327 Campus Dr, Stanford, CA 94305, USA. Electronic address: sjdixon@stanford.edu.

Abstract

Mammalian cells can die by apoptosis or by one of several non-apoptotic mechanisms, such as ferroptosis. Here, we present a protocol to distinguish ferroptosis from other cell death mechanisms in cultured cells. We describe steps for seeding cells, administering mechanism-specific cell death inducers and inhibitors, and measuring cell death and viability. We then detail the use of molecular markers to verify mechanisms of cell death. This protocol can be used to identify and distinguish ferroptosis in 2D and 3D cultures. For complete details on the use and execution of this protocol, please refer to Ko, et al. (2019),¹ Magtanong, et al. (2022),² and Armenta, et al. (2022).³.

Keywords: Cancer; Cell biology; Cell culture; Cell-based assays.

MeSH terms

Animals
Apoptosis
Cell Death
Cells, Cultured
Ferroptosis*
Mammals