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Fig. 6. Osteogenic differentiation ability of both hMSCs and hmNSCs. Both cell types were differentiated after 6-10 passages using the osteoblast differentiation protocol for 10 days. (A) Differentiated hMSCs (upper panel) and hmNSCs (lower panel) were stained for the osteogenic marker alkaline phosphatase (AP). Bar, 50 µm. (B) Quantification of 10-day cultures of hMSCs and hmNSCs differentiated into osteoblasts under normal atmospheric oxygen levels routinely used for osteoblast cultures (21%) and reduced atmospheric oxygen levels used in our neural culture system (3%). Data shown are mean±s.e.m. from at least three independent cell preparations. *, P<0.05; **, P<0.01 when compared to the percentage of AP+ cells in hMSCs. (C) Expression of osteogenic and mesodermal marker genes as well as the transcription factor c-fos in hMSCs and hmNSCs after osteogenic differentiation under 21% (lanes 1, 4) and 3% (lanes 2, 3) atmospheric oxygen levels. Semiquantitative RT-PCR analysis of AP, runt-related transcription factor 2 (RUNX2), SOX9 and the transcription factor c-fos, as well as GAPDH (housekeeping gene) was performed in hmNSC (lanes 1 and 2) and hMSC (lanes 3 and 4).





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