Fig. 5

Clonal isolation of the PDGFRα⁺/CD90⁺ subpopulation. a Gating strategy for the single-cell isolation of PDGFRα⁺/CD90⁺ cells. Cells were initially gated on FSC and SSC followed by CD45⁻/Ter119⁻ cells (upper). Gated Sca-1⁺ cells among CD45⁻/Ter119⁻ cells (lower left) are then subdivided into the four CD90/PDGFRα subpopulations (lower right) where percentages in each quadrant are indicated. b Distribution of subpopulations in fresh cBM. c Bright-field images of expanded cells of the indicated subpopulation after 10 days of culturing. d CFU-F frequencies of Sca-1⁺ cells in fBM (left), cBM (middle) and PDGFRα⁺CD90⁺Sca-1⁺ cBM (right) cells. Expanded PDGFRα⁺/CD90⁺ cells after 7 days in e adipogenic and f osteogenic differentiation media. First column = 4× magnification (image bar = 200 μm) and second column = 10× magnification (image bar = 100 μm). g Survival curve for PDGFRα⁺/CD90⁺ colonies during in-vitro expansion. h Representative histogram of CFSE staining among gated T cells of CD3/CD28 stimulated splenocytes with and without addition of cloned PDGFRα⁺/CD90⁺ cells (white, solid line, stimulated splenocytes; tinted, dashed line, unstimulated splenocytes). Bar = 200 μm. i Percentage immunosuppression among different subsets of T lymphocytes. cBM compact bone marrow, fBM flushed bone marrow, MSC mesenchymal stromal cell, PDGFRα platelet-derived growth factor receptor alpha