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

Deshan Ren; Wei Liu; Shuai Ding; Yan Li

doi:10.1016/j.xpro.2022.101217

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

STAR Protoc. 2022 Mar 3;3(1):101217. doi: 10.1016/j.xpro.2022.101217. eCollection 2022 Mar 18.

Authors

Deshan Ren¹, Wei Liu¹, Shuai Ding², Yan Li¹

Affiliations

¹ MOE Key Laboratory of Model Animals for Disease Study, MOE Engineering Research Center of Protein and Peptide Medicine, The State Key Laboratory of Pharmaceutical Biotechnology, Chemistry and Biomedicine Innovation Center, Model Animal Research Center, Medical School of Nanjing University, Nanjing 210061, China.
² Department of Rheumatology and Immunology, Nanjing Drum Tower Hospital, The Affiliated Hospital of Nanjing University Medical School, Nanjing, China.

Abstract

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).

Keywords: Biotechnology and bioengineering; Cell isolation; Flow Cytometry/Mass Cytometry; Immunology; Model Organisms.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Flow Cytometry / methods
Hematopoiesis
Humans
Hydrodynamics*
Immune System*
Mice