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

Figure 6

Influence of passaging on hBMSC differentiation. hBMSC in passage 1 until passage 9 were plated in BM (basic medium, white columns) onto TCPS (= P2–P10 for the analyses). From day 4 after plating, the cells were cultured either in BM or in OM/D (osteogenic differentiation medium, black columns). At day 15 after plating, the cells were analyzed for expression of oct-4 (a), cbfa1 (b), pthr (d), bglap (e), and pparγ (f), and for TNAP activity (c) as described above. 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 passage 2 value in BM = 1. Small digits in the black columns (OM/D) indicate the inducibility of the parameter in each passage as quotient OM/D : BM. Significant differences of OM/D versus BM for each passage were analyzed by two-way ANOVA with Bonferroni’s post-test and indicated (), .

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