Fig. 3

Hypoxic culturing increase the differentiation potential of hBMMSCs. Sample-matched hBMMSCs cultured in atmospheric or hypoxic conditions for 3 passages from the time of isolation were tested for their ability to differentiate into osteoblast (a, b), adipocyte (c, d) and chondrocyte (e). a, c, e hBMMSCs were able to undergo tri-lineage differentiation regardless of oxygen pressure, but demonstrated increased chondrocyte specific differentiation under hypoxia, specifically COL2A1 and ACAN. Relative gene expression by qPCR. a Expression of CBFA1 and osteocalcin (OC) 15 days after differentiation in osteogenic medium. c Expression of CEBPa and FABP4 15 days after differentiation in adipogenic medium. e Expression of Sox9, COL1A1, COL2A1 and ACAN 21 days after differentiation in chondrogenic medium. Data represent mean ± SEM, n = 3 independent sample-matched hBMMSCs. Two-tailed paired Student’s t-test *p value ≤ 0.05. b, d Representative micrographs of undifferentiated hBMMSCs (day 0) and hBMMSCs 15 days after differentiation into osteoblasts (b) or adipocytes (d)