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

• Published in: Nature communications 2021-06, Vol.12 (1), p.3568-3568, Article 3568
• Main Authors: Ochi, Kiyosumi, Morita, Maiko, Wilkinson, Adam C., Iwama, Atsushi, Yamazaki, Satoshi
• Format: Article
• Language: English
• Subjects: 13/100; 13/31; 49/1; 631/1647/1511; 631/1647/2300; 631/250; 631/250/1904; 631/532/1542; Animal models; Animals; Bone marrow; Bone Marrow - metabolism; Bone Marrow Cells; Bone Marrow Transplantation; Cell culture; Cells (biology); Chimera - metabolism; Chimeras; Conditioning; Culture techniques; Doxycycline; Enrichment; Flow cytometry; Fluorescence; Gene Transfer Techniques; Genetic Engineering; Genetic Therapy; Hematopoiesis; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Hematopoietic Stem Cells - metabolism; Humanities and Social Sciences; Immune system; Immunology; Irradiation; Male; Mice; Mice, Inbred C57BL; multidisciplinary; Osteoprogenitor cells; Polyvinyl alcohol; Progenitor cells; Radiation; Science; Science (multidisciplinary); Transplantation; Transplantation Chimera
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-021-23763-z

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. Bone marrow chimaeric mice are a valuable tool in research, but require myeloablative conditioning. Here the authors demonstrate efficient FACS-free enrichment of haematopoietic stem and progenitor cells for transplantation into unconditioned recipient mice, as well as for genetic engineering using polyvinyl alcohol based media.