Assessment of Ferroptosis in Hematopoietic Stem and Progenitor Cells

Hematopoietic stem cells (HSCs) are critical for maintaining hematopoiesis throughout life by utilizing their self-renewing and multipotent capabilities. Ferroptosis is a type of cell death characterized by the iron-dependent accumulation of lipid peroxides, and it is involved in multiple physiological and pathological conditions. Recent studies have highlighted the important role of ferroptosis in the functional maintenance of HSCs. Here, we describe our current protocols for accessing ferroptosis in hematopoietic stem and progenitor cells (HSPCs) both in vivo and in vitro. We introduce procedures for measuring total reactive oxygen species (ROS) and lipid ROS in HSPCs, as well as analyzing cell number, cell viability, and cell cycle profiles. This protocol provides a useful approach for characterizing the status of ferroptosis and its related parameters in HSPCs and more broadly, for studying the outcomes of ferroptosis on hematopoiesis.