Human Bone Marrow Stromal Cells: A Reliable, Challenging Tool for In Vitro Osteogenesis and Bone Tissue Engineering Approaches

Figure 5

Influence of culture time and dexamethasone on hBMSC differentiation. hBMSC in passage 1 were plated in BM (basic medium) onto TCPS (= P2). From day 4 after plating, the cells were cultured either in BM (open circles) or in OM/D (osteogenic differentiation medium; black circles). At indicated time points (days 1, 2, and 4 in BM, and days 8, 11, 15, 18, 22, and 25 in both BM and OM/D) the cells were analyzed for the expression of octamer binding transcription factor 4 (oct-4, (a)), core-binding factor subunit alpha-1 (cbfa1, (b)), parathyroid hormone receptor (pthr, (d)), bone gla protein (bglap, (e)), and peroxisome proliferator-activated protein γ (pparγ, (f)). Relative gene expression (gene of interest) was assessed by real time PCR and normalized to the expression of beta-actin as house-keeping gene by comparative quantitation analysis setting day 1 in BM = 1. TNAP activity (c) was determined as described above. Significant differences of OM/D versus BM were analyzed for each time point with -test and were indicated (), .

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