Fig. 3

Differentiation potential of older donor MSCs as a function of CD264⁺ cell content. P7 MSCs from older donors, 45–60 years old, were cultured for 21 days in either osteogenic or adipogenic medium. a Representative low and high CD264 cultures stained with Alizarin Red S to detect mineralization during osteogenesis and AdipoRed to detect lipid accumulation during adipogenesis. Negative control cultures maintained in growth medium. Scale bars = 100 μm. b,c Absorbance from extracted Alizarin Red S (b) and fluorescence from AdipoRed (c), respectively, are reported per microgram DNA and standardized relative to the negative control. MSCs were harvested from male (circle) and female (triangle) donors. Data are reported as mean ± SEM for n = 3 biological replicates. Nonparametric linear regression lines in panels b and c shown for n = 6 donor cultures