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Mesenchymal precursor cells (MPCs) are isolated from human bone marrow aspirates by density gradient separation, over Ficoll, followed by STRO-3-based immunoselective magnetic activated cell sorting (MACS); cultures are then expanded. STRO-3 is a monoclonal antibody, which reacts with an epitope of non-specific, human tissue alkaline phosphatase. Stro3 binds to the immunizing stromal cells, but lacks reactivity with peripheral blood-derived mononuclear cells.
Culture-expanded MPCs demonstrate increased gene expression of a range of stem cell markers, when compared to mesenchymal stem cells (MSCs), and exhibit a greater capacity to undergo osteogenic, adipogenic and chondrogenic differentiation.
STRO-3-selected MPCs were plated as single-cell suspensions at a density of 8x103 cells per cm2, in αMEM supplemented with FCS (10%), L-ascorbate-2-phosphate (100 µM), β-mercaptoethanol (50 µM) and Primocin (100 µg/ml).