Phenotype of MPCs.
CD45−GlyA− cells were plated on FN in expansion medium with 10 ng/mL EGF and 10 ng/mL PDGF-BB, and passaged for 15 to 40 cell doublings. Cells were harvested and labeled with antibodies against CD10, CD13, CD31, CD34, CD36, CD38, CD44, CD49b, CD50, CD62P, CDw90, CD106, CD117, H1P12, IB10, KDR, Flt1, HLA-DR, class I HLA, and β2-microglobulin or control IgGs, as indicated and analyzed by FACS. Plots show isotype control IgG-staining profile (thin line) versus specific antibody staining profile (thick line). A representative example of more than 5 experiments is shown. (A) Cells were cultured with 2% FCS at a cell density between 2 and 8 × 103/cm2 for 15 cell doublings. (B) cells were cultured with 2% FCS at a cell density between 2 and 8 × 103/cm2 for 40 cell doublings. (C) Cells were cultured without FCS but with 10 ng/mL IGF-1 at a cell density between 2 and 8 × 103/cm2 for 15 cell doublings. (D) Cells were plated with 10% FCS, at a cell density between 2 and 8 × 103/cm2 for 15 cell doublings. (E) Cells were plated with 2% FCS at a cell density between greater than 104/cm2 for 15 cell doublings.
