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Identification of Nonhuman Primate Hematopoietic Stem and Progenitor Cells

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Hematopoietic Stem Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2567))

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Abstract

The preclinical development of hematopoietic stem cell (HSC) gene therapy/editing and transplantation protocols is frequently performed in large animal models such as nonhuman primates (NHPs). Similarity in physiology, size, and life expectation as well as cross-reactivity of most reagents and medications allows for the development of treatment strategies with rapid translation to clinical applications. Especially after the adverse events of HSC gene therapy observed in the late 1990s, the ability to perform autologous transplants and follow the animals long-term make the NHP a very attractive model to test the efficiency, feasibility, and safety of new HSC-mediated gene-transfer/editing and transplantation approaches.

This protocol describes a method to phenotypically characterize functionally distinct NHP HSPC subsets within specimens or stem cell products from three different NHP species. Procedures are based on the flow-cytometric assessment of cell surface markers that are cross-reactive in between human and NHP to allow for immediate clinical translation. This protocol has been successfully used for the quality control of enriched, cultured, and gene-modified NHP CD34+ hematopoietic stem and progenitor cells (HSPCs) as well as sort-purified CD34 subsets for transplantation in the pig-tailed, cynomolgus, and rhesus macaque. It further allows the longitudinal assessment of primary specimens taken during the long-term follow-up post-transplantation in order to monitor homing, engraftment, and reconstitution of the bone marrow stem cell compartment.

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Author information

Authors and Affiliations

  1. Stem Cell and Gene Therapy Program, Fred Hutchinson Cancer Research Center, Seattle, WA, USA

    Stefan Radtke & Hans-Peter Kiem

  2. Department of Medicine, University of Washington School of Medicine, Seattle, WA, USA

    Hans-Peter Kiem

  3. Department of Pathology, University of Washington School of Medicine, Seattle, WA, USA

    Hans-Peter Kiem

Authors
  1. Stefan Radtke
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  2. Hans-Peter Kiem
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Correspondence to Stefan Radtke .

Editor information

Editors and Affiliations

  1. Department of Microbiology & Immunology and Department of Medicine/Hematology Oncology, Indiana University School of Medicine, Indianapolis, IN, USA

    Louis M. Pelus

  2. Moderna Therapeutics, Cambridge, MA, USA

    Jonathan Hoggatt

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Radtke, S., Kiem, HP. (2023). Identification of Nonhuman Primate Hematopoietic Stem and Progenitor Cells. In: Pelus, L.M., Hoggatt, J. (eds) Hematopoietic Stem Cells. Methods in Molecular Biology, vol 2567. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2679-5_6

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  • DOI: https://doi.org/10.1007/978-1-0716-2679-5_6

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2678-8

  • Online ISBN: 978-1-0716-2679-5

  • eBook Packages: Springer Protocols

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