Non-conditioned bone marrow chimeric mouse generation using culture-based enrichment of hematopoietic stem and progenitor cells

Kiyosumi Ochi, Maiko Morita, Adam C. Wilkinson, Atsushi Iwama and Satoshi Yamazaki ()
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Kiyosumi Ochi: The University of Tokyo
Maiko Morita: The University of Tokyo
Adam C. Wilkinson: University of Oxford, Radcliffe Department of Medicine, University of Oxford
Atsushi Iwama: The University of Tokyo
Satoshi Yamazaki: The University of Tokyo

Nature Communications, 2021, vol. 12, issue 1, 1-10

Abstract: Abstract Bone marrow (BM) chimeric mice are a valuable tool in the field of immunology, with the genetic manipulation of donor cells widely used to study gene function under physiological and pathological settings. To date, however, BM chimera protocols require myeloablative conditioning of recipient mice, which dramatically alters steady-state hematopoiesis. Additionally, most protocols use fluorescence-activated cell sorting (FACS) of hematopoietic stem/progenitor cells (HSPCs) for ex vivo genetic manipulation. Here, we describe our development of cell culture techniques for the enrichment of functional HSPCs from mouse BM without the use of FACS purification. Furthermore, the large number of HSPCs derived from these cultures generate BM chimeric mice without irradiation. These HSPC cultures can also be genetically manipulated by viral transduction, to allow for doxycycline-inducible transgene expression in donor-derived immune cells within non-conditioned immunocompetent recipients. This technique is therefore expected to overcome current limitations in mouse transplantation models.

Date: 2021
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DOI: 10.1038/s41467-021-23763-z

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