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

Nature Communications
Kiyosumi OchiSatoshi Yamazaki

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.

References

Nov 23, 2000·Proceedings of the National Academy of Sciences of the United States of America·S HatadaO Smithies
Sep 24, 2002·Nature Immunology·Simon FillatreauStephen M Anderton
Jan 12, 2011·Proceedings of the National Academy of Sciences of the United States of America·Xu CaoFrank J Frassica
May 28, 2011·Methods in Molecular Biology·Lara RossiMargaret A Goodell
Feb 13, 2013·Methods in Molecular Biology·Kyle RectorGary Van Zant
Feb 11, 2014·Methods in Molecular Biology·Marcia Regina Machein, Karl H Plate
Apr 14, 2015·Journal of Visualized Experiments : JoVE·Kyle PeakeCharles Krieger
May 24, 2016·Current Opinion in Hematology·Katrin Busch, Hans-Reimer Rodewald
Aug 7, 2018·Frontiers in Oncology·Ursa Lampreht TratarMaja Cemazar
Jan 10, 2020·Nature Protocols·Adam C WilkinsonSatoshi Yamazaki

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Citations

Aug 21, 2021·Transplantation·Prince Singh, Xunrong Luo

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