Analysis of osteogenic and adipogenic potential.

Using Alizarin S red staining, Calcium deposits could be detected in P13 MCs (B) and P21 MSCs (D), indicating osteogenic differentiation, while cells under control conditions failed to exhibit the deposits (A, C). Fat droplet accumulation could be detected in P5 MCs (F) and, slightly more pronounced in MSCs (H). Control conditions showed no generation of fat droplets (E, G). Expression analysis by qPCR revealed that the osteogenic marker Sparc was up-regulated in the earlier passages (I), with a significant 6-fold change in P13 MCs. A significant 3.7-fold increase in expression was observed in P5 MCs for the adipogenic gene PPARγ (K). A Student’s t-test was used to calculate significance. Scale bars are 30 μM.