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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Ildstad ST, Shirwan H, Leventhal J (2011) Is durable macrochimerism key to achieving clinical transplantation tolerance? Curr Opin Organ Transplant 16:343–344
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
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
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
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
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
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
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
Kiel MJ, Yilmaz OH, Morrison SJ (2008) CD150 cells are transiently reconstituting multipotent progenitors with little or no stem cell activity. Blood 111:4413–4414
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
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
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
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.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Menendez-Gonzalez, J.B., Saleh, L., Feng, G.J., Rodrigues, N.P. (2019). Isolation of Murine Hematopoietic Stem Cells. In: Boyd, A. (eds) Immunological Tolerance. Methods in Molecular Biology, vol 1899. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8938-6_2
Download citation
DOI: https://doi.org/10.1007/978-1-4939-8938-6_2
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-8936-2
Online ISBN: 978-1-4939-8938-6
eBook Packages: Springer Protocols