Protocol to construct humanized mice with adult CD34 + hematopoietic stem and progenitor cells.

Humanized mice, defined as mice with human immune systems, have become an emerging model to study human hematopoiesis, infectious disease, and cancer. Here, we describe the techniques to generate humanized NSGF6 mice using adult human CD34 ⁺ hematopoietic stem and progenitor cells (HSPCs). We describe steps for constructing and monitoring the engraftment of humanized mice. We then detail procedures for tissue processing and immunophenotyping by flow cytometry to evaluate the multilineage hematopoietic differentiation. For complete details on the use and execution of this protocol, please refer to Yu et al. ¹ .
Competing Interests: Declaration of interests C.I.Y., R.M., J.B., and K.P. filed a patent on novel humanized mouse models via genetic editing of NSG mouse. K.P. is a stockholder in Cue Biopharma and Guardian Bio, scientific advisor to Cue Biopharma and Guardian Bio, and co-founder of Guardian Bio. K.P. declares unrelated funding support from Guardian Bio (current) and Merck (past). J.B. is associated with Immunai.
(Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)