Isolation, Differentiation, and Characterization of Human Bone Marrow Stromal Stem Cells In Vitro and In Vivo.

In this chapter, we describe techniques for the isolation, enrichment, and characterization of skeletal stem cells from human bone marrow. The methods for enriching STRO-1 and STRO-4 positive cells using magnetic activated cell sorting are detailed, as are the techniques for establishing and characterising osteogenic, adipogenic, and chondrogenic cultures derived from these cells. Finally, we present methods for studying the ability of these enriched skeletal stem and progenitor populations to produce bone in vivo using diffusion chambers following subcutaneous implantation, over 4-6 weeks.