Isolation and culture of mesenchymal stem cells from mouse compact bone

The bone marrow (BM) of numerous species, including rodents and man, contains a rare population of cells termed marrow stromal cells or mesenchymal stem cells (MSC). Given the ability of these cells to differentiate into cells of the osteogenic, chondrogenic and adipogenic lineages, there is considerable interest in utilizing MSCs in a broad repertoire of cell-based therapies for the treatment of human disease. Before such potential therapies can be realized, a preclinical animal model in which to test and refine strategies utilizing MSC is required. Here we describe methods for the isolation of a highly enriched population of MSC from mouse cortical/compact bone (CB), quantitation using the colony forming unit-fibroblast assay (CFU-F) and in vitro expansion. These cells are both multipotent and capable of extensive in vitro expansion and thus represent an ideal cellular source to explore both the biological properties of MSC as well as their potential efficacy in a variety of cellular therapies.