Summary
Hematopoietic stem cells (HSC) are rare with estimated frequencies of 1 in 10,000 bone marrow cells and 1 in every 100,000 blood cells. The most important characteristic of HSC is their capacity to provide complete restoration of all blood cell lineages after bone marrow ablation. Therefore they are considered as the ideal targets for various clinical applications including stem cell transplantation and gene therapy. In adult mice and men, the main stem cell source is the bone marrow. For clinical applications HSC derived from umbilical cord blood (UCB) and G-CSF mobilized peripheral blood (PB) have been demonstrated to have several advantages compared to bone marrow; therefore, they are slowly replacing BM as alternative source of stem cells. The mouse is the model organism of choice for immunological and hematological research; therefore, studies of murine HSC are an important research topic. Here we described the most often used protocols and methods to isolate human and mouse HSC to high purity.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Kondo, M., Wagers, A.J., Manz, M.G., Prohaska, S.S., Scherer, D.C., Beilhack, G.F., Shizuru, J.A., Weissman, I.L. (2003) Biology of hematopoietic stem cells and progenitors: implications for clinical application. Annu Rev Immunol. 21, 759 – 806
Coulombel, L. (2004) Identification of hemat-opoietic stem/progenitor cells: strength and drawbacks of functional assays. Oncogene. 23, 7210 – 7222
Gangenahalli, G.U., Singh, V.K., Verma, Y.K., Gupta, P., Sharma, R.K., Chandra, R., Luthra, P.M. (2006) Hematopoietic stem cell antigen CD34: role in adhesion or homing. Stem Cells Dev. 15, 305 – 313
Buhring, H.J., Asenbauer, B., Katrilaka, K., Hummel, G., and Busch, F.W. (1989) Sequential expression of CD34 and CD33 antigens on myeloid colony-forming cells. Eur J Haematol. 42, 143 – 149
Ng, Y. Y., Bloem, A.C., van Kessel, B., Lokhorst, H., Logtenberg, T., Staal, F.J. (2002) Selective in vitro expansion and efficient retroviral trans-duction of human CD34+ CD38 haematopoietic stem cells. Br J Haematol. 117, 226 – 237
Ng, Y.Y., van Kessel, B., Lokhorst, H.M., Baert, M.R., van den Burg, C.M., Bloem, A.C., Staal, F.J. (2004) Gene-expression profiling of CD34+ cells from various hemat-opoietic stem-cell sources reveals functional differences in stem-cell activity. J Leukoc Biol. 75, 314 – 323
Pike-Overzet, K., de Ridder, D., Weerkamp, F., Baert, M.R., Verstegen, M.M., Brugman, M.H., Howe, S.J., Reinders, M.J., Thrasher, A.J., Wagemaker, G., van Dongen, J.J., Staal, F.J. (2007) Ectopic retroviral expression of LMO2, but not IL2Rgamma, blocks human T-cell development from CD34+ cells: implications for leukemogenesis in gene therapy. Leukemia. 21, 754 – 763
Lagasse E., Connors H., Al-Dhalimy M., Reitsma M., Dohse, M., Osborne, L., Wang, L., Finegold, M., Weissman, I.L., Grompe, M. (2000) Purified hematopoietic stem cells can differentiate into hepatocytes in vivo. Nat Med. 6, 1229 – 1234
BD Biosciences (2006). BD FACSAria User's Guide
Acknowledgments
The authors would like to thank Tiago Luis and Sjanneke Heuv-elmans for techincal assistance.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Ng, Y.Y., Baert, M.R., de Haas, E.F., Pike-Overzet, K., Staal, F.J. (2009). Isolation of Human and Mouse Hematopoietic Stem Cells. In: Baum, C. (eds) Genetic Modification of Hematopoietic Stem Cells. Methods In Molecular Biology™, vol 506. Humana Press. https://doi.org/10.1007/978-1-59745-409-4_2
Download citation
DOI: https://doi.org/10.1007/978-1-59745-409-4_2
Publisher Name: Humana Press
Print ISBN: 978-1-58829-980-2
Online ISBN: 978-1-59745-409-4
eBook Packages: Springer Protocols