Isolation of human and murine hematopoietic stem cells for DNA damage and DNA repair assays

Summary Hematopoietic stem and progenitor cells (HSPCs) reside in the bone marrow and supply blood cells. Efficient methods for isolation of HSPCs are required. Here, we present protocols for the isolation of human and murine HSPCs using manual and FACS-assisted techniques. Isolated HSPCs can be used for downstream applications, including colony forming unit assays and DNA damage and repair assays. For complete details on the use and execution of this protocol, please refer to Rodríguez et al. (2021a) and (2021b).
Graphical abstract
Highlights • Detailed protocol for isolating human and mouse hematopoietic stem cells • Procedures for plating and quantification of hematopoietic colony forming unit assay • Protocol for assessing DNA damage using the comet assay in hematopoietic stem cells • Protocol for assessing DNA damage using immunofluorescence in hematopoietic stem cells
Hematopoietic stem and progenitor cells (HSPCs) reside in the bone marrow and supply blood cells. Efficient methods for isolation of HSPCs are required. Here, we present protocols for the isolation of human and murine HSPCs using manual and FACS-assisted techniques. Isolated HSPCs can be used for downstream applications, including colony forming unit assays and DNA damage and repair assays.