Abstract
Current bone disease therapy with bone marrow-derived mesenchymal stromal cells (MSC) is hampered by low efficiency. Advanced allogeneic studies on well-established mouse genetic and disease models are hindered by difficulties in isolating murine MSC (mMSC). And mMSC prepared from different laboratories exhibit significant heterogeneity. Hence, this study aimed to identify and isolate a sub-population of mMSC at an early passage number with high osteogenic potential. Enrichment of mMSC was achieved by 1-hr silica incubation and negative selection. Approximately 96% of these cells synthesized osteocalcin after 28 days of osteogenic induction in vitro, and displayed a complete dynamic alteration of alkaline phosphatase (ALP) activity with increasing osteogenic maturation and strong mineralization. Moreover, the cells displayed uniform and stable surface molecular profile, long-term survival, fast proliferation in vitro with maintenance of normal karyotype and distinct immunological properties. CD73 was found to be expressed exclusively in osteogenesis but not in adipogenesis. These cells also retained high osteogenic potential upon allogeneic transplantation in an ectopic site by the detection of bone-specific ALP, osteopontin, osteocalcin and local mineralization as early as 12 days after implantation. Hence, these cells may provide a useful source for improving current strategies in bone regenerative therapy, and for characterizing markers defining the putative MSC population.