Protocol for generating human immune system mice and hydrodynamic injection to analyze human hematopoiesis in vivo

Human immune system (HIS) mice provide a valuable platform to investigate and modulate human hematopoiesis development in vivo. Here, we describe detailed protocols for the construction of HIS mice, modulation of human hematopoiesis in vivo using hydrodynamic injection of plasmids encoding cytokines of interest, and flow cytometry analysis of humanization levels and human immune subsets. This approach can be easily applied to screen or verify factors that regulate human hematopoiesis and immune system. For complete details on the use and execution of this protocol, please refer to Cardoso et al. (2021) and Li et al. (2017).