Advertisement

Isolation of Murine Hematopoietic Stem Cells

  • Juan Bautista Menendez-Gonzalez
  • Lubaid Saleh
  • Gui Jie Feng
  • Neil P. RodriguesEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1899)

Abstract

Bone marrow resident hematopoietic stem cells (HSCs) are responsible for the lifetime generation of the wide profusion of blood and immune cell types found in the body. In addition, therapeutically, in the context of bone marrow transplantation, HSCs have been successfully deployed to restore normal blood-forming capacity in patients being treated with high-dose chemotherapy for hematologic malignancies. The known ability of bone marrow transplantation to either restore or reset the immune system and to engender immune tolerance has suggested that HSCs may be applied therapeutically for a wider range of clinical conditions, including immunological/autoimmune disorders and allogeneic organ transplantation. Herein, we describe a flow-cytometry-based method to isolate mouse HSCs for continued experimental investigation into such therapeutic uses.

Key words

HSCs Tolerance Transplantation Flow cytometry Cell sorting 

Notes

Acknowledgments

This work was supported by the Life Science Research Network Wales, an initiative funded through the Welsh Government’s Ser Cymru program, Leukaemia Cancer Society, Leukaemia Research Appeal for Wales and the Lady Tata Memorial Trust. JBMG was funded by a Ser Cymru Richard Whipp Studentship.

References

  1. 1.
    Ildstad ST, Shirwan H, Leventhal J (2011) Is durable macrochimerism key to achieving clinical transplantation tolerance? Curr Opin Organ Transplant 16:343–344PubMedCrossRefGoogle Scholar
  2. 2.
    Leventhal J, Abecassis M, Miller J et al (2012) Chimerism and tolerance without GVHD or engraftment syndrome in HLA-mismatched combined kidney and hematopoietic stem cell transplantation. Sci Transl Med 4:124ra28. https://doi.org/10.1126/scitranslmed.3003509 PubMedPubMedCentralCrossRefGoogle Scholar
  3. 3.
    Shizuru JA, Weissman IL, Kernoff R et al (2000) Purified hematopoietic stem cell grafts induce tolerance to alloantigens and can mediate positive and negative T cell selection. Proc Natl Acad Sci U S A 97:9555–9560. https://doi.org/10.1073/pnas.170279297 PubMedPubMedCentralCrossRefGoogle Scholar
  4. 4.
    Zheng J, Umikawa M, Zhang S et al (2011) Ex vivo expanded hematopoietic stem cells overcome the MHC barrier in allogeneic transplantation. Cell Stem Cell 9:119–130. https://doi.org/10.1016/j.stem.2011.06.003 PubMedPubMedCentralCrossRefGoogle Scholar
  5. 5.
    Fujisaki J, Wu J, Carlson AL et al (2011) In vivo imaging of T reg cells providing immune privilege to the haematopoietic stem-cell niche. Nature 474:216–220. https://doi.org/10.1038/nature10160 PubMedPubMedCentralCrossRefGoogle Scholar
  6. 6.
    Sullivan KM, Goldmuntz EA, Keyes-Elstein L et al (2018) Myeloablative autologous stem-cell transplantation for severe scleroderma. N Engl J Med 378:35–47. https://doi.org/10.21430/M3SM4LTLH PubMedPubMedCentralCrossRefGoogle Scholar
  7. 7.
    Forsberg EC, Prohaska SS, Katzman S et al (2005) Differential expression of novel potential regulators in hematopoietic stem cells. PLoS Genet 1:e28. https://doi.org/10.1371/journal.pgen.0010028 PubMedPubMedCentralCrossRefGoogle Scholar
  8. 8.
    Kiel MJ, Yilmaz ÖH, Iwashita T et al (2005) SLAM family receptors distinguish hematopoietic stem and progenitor cells and reveal endothelial niches for stem cells. Cell 121:1109–1121. https://doi.org/10.1016/j.cell.2005.05.026 PubMedCrossRefGoogle Scholar
  9. 9.
    Kiel MJ, Yilmaz OH, Morrison SJ (2008) CD150 cells are transiently reconstituting multipotent progenitors with little or no stem cell activity. Blood 111:4413–4414PubMedPubMedCentralCrossRefGoogle Scholar
  10. 10.
    Kim I, He S, Yilmaz ÖH et al (2006) Enhanced purification of fetal liver hematopoietic stem cells using SLAM family receptors. Blood 108:737–744. https://doi.org/10.1182/blood-2005-10-4135 PubMedPubMedCentralCrossRefGoogle Scholar
  11. 11.
    Yilmaz ÖH, Kiel MJ, Morrison SJ (2006) SLAM family markers are conserved among hematopoietic stem cells from old and reconstituted mice and markedly increase their purity. Blood 107:924–930. https://doi.org/10.1182/blood-2005-05-2140 PubMedPubMedCentralCrossRefGoogle Scholar
  12. 12.
    Oguro H, Ding L, Morrison SJ (2013) SLAM family markers resolve functionally distinct subpopulations of hematopoietic stem cells and multipotent progenitors. Cell Stem Cell 13:102–116. https://doi.org/10.1016/j.stem.2013.05.014 PubMedPubMedCentralCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Juan Bautista Menendez-Gonzalez
    • 1
  • Lubaid Saleh
    • 1
  • Gui Jie Feng
    • 1
  • Neil P. Rodrigues
    • 1
    Email author
  1. 1.European Cancer Stem Cell Research InstituteCardiff University, School of BiosciencesCardiffUK

Personalised recommendations