140,000 MSC per well were seeded in 6-well plates in 1% FCS/DMEM and allowed to adhere overnight. After incubation for 3 weeks as indicated, cells were fixed with ice-cold methanol for 30 min at − 20 °C and air dried. 5% Alizarin (1,2-dihydroxyanthraquinone, Sigma) in 0.1 M boric acid buffer (pH 4) was filtered and applied to the cells for 1 h at room temperature. After several wash steps with PBS (pH 6.0), wells were dried and micrographs were taken on a Zeiss Axiovert 40 CFL using a Canon PowerShot A649.

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