**3.1** *In vitro* **osteogenesis of human BM-MSCs**

Osteogenic medium: MSC growth medium supplemented with 50 μM ascorbic acid phosphate (AsAP) (Wako Chemicals USA, Richmond, VA), 0.1 μM dexamethasone (Sigma-Aldrich, St Louis, MO), and 10 mM β-glycerol phosphate (Sigma-Aldrich, St Louis, MO).

#### **Figure 1.**

*Osteogenic culture of human BM-MSCs. Representative images (10×) of human BM-MSC osteogenesis on day 12 (A), day 17 (B), and day 26 during osteogenic culture.*

Several osteogenic markers, such as alkaline phosphatase (ALP), leptin receptor, and cathepsin K, are used as the indicator of early osteogenesis [28], calcium deposition as the indicator of late-stage osteogenesis [28, 29]. ALP activity and calcium deposition are exemplified to assess osteogenic differentiation of human MSCs from passage 4 (**Figure 1**).
