Abstract
In this chapter, we describe techniques for the isolation and characterization of skeletal stem cells from human bone marrow. The methods for enrichment of STRO-1 positive cells using magnetic activated cell sorting are described and we also cover techniques for establishing and characterising osteogenic, adipogenic, and chondrogenic cultures from these cells. Finally, we present methods for studying the ability of these cells to produce bone in vivo using diffusion chambers which have been implanted subcutaneously in mice.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Simmons, P., and Torok-Storb, B. (1991) Identification of stromal cell precursors in human bone marrow by a novel monoclonal antibody, STRO-1. Blood 78, 55–62.
Stewart, K., Walsh, S., Screen, J., Jefferiss, C. M., Chainey, J., Jordan, G. R., and Beresford, J. N. (1999) Further characterisation of cells expressing STRO-1in cultures of adult human bone marrow stromal cells. J. Bone Miner. Res. 14, 1345–1356.
Gundle, R., Joyner, C. J., and Triffitt, J. T. (1995) Human bone tissue formation in diffusion chamber culture in vivo by bone-derived cells and marrow stromal fibroblastic cells. Bone 16, 597–601.
Oreffo, R. O., and Triffitt, J. T. (1999) In vitro and in vivo methods to determine the interactions of osteogenic cells with biomaterials. J. Mater. Sci. Mater. Med. 10, 607–611.
Bolland, B. J., Kanczler, J. M., Dunlop, D. G., and Oreffo, R. O. (2008) Development of in vivo mCT evaluation of neovascularisation in tissue engineered bone constructs. Bone 43, 195–202.
Bolland, B. J., Kanczler, J. M., Ginty, P. J., Howdle, S. M., Shakesheff, K. M., Dunlop, D. G., and Oreffo, R. O. (2008) The application of human bone marrow stromal cells and poly(dl-lactic acid) as a biological bone graft extender in impaction bone grafting. Biomaterials 29, 3221–3227.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Tare, R.S., Mitchell, P.D., Kanczler, J., Oreffo, R.O.C. (2012). Isolation, Differentiation, and Characterisation of Skeletal Stem Cells from Human Bone Marrow In Vitro and In Vivo. In: Helfrich, M., Ralston, S. (eds) Bone Research Protocols. Methods in Molecular Biology, vol 816. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-415-5_7
Download citation
DOI: https://doi.org/10.1007/978-1-61779-415-5_7
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-414-8
Online ISBN: 978-1-61779-415-5
eBook Packages: Springer Protocols