Fig. 2

In vitro-cultured hBM-MSCs respond to rhBMP-2 treatment. A In vitro cell culture growth kinetics of hBM-MSCs. (Cells from 9 donors were included). B hBM-MSCs appearance at early cell culture (10 days) and late cell culture (70 Days) points. C ALP staining (red) of early hBM-MSCs cultured for 7 days with rhBMP-2. D Multiple parameters of hBM-MSCs treated for 7 days at different time points. The data are presented relative to the number of days of culture for the hBM-MSCs. Colorimetric ALP activity was measured as an indicator of an early differentiation marker, ethidium homodimer DNA was used as an indicator of a proliferation marker, and MTT mitochondrial activity was used as an indicator of cell metabolic activity. All the data are presented normalized to their own control on the day of the start of the assay. The first assay was started on day 14 after starting the cell culture. n = 3 for each experimental point. The data shown correspond to donor D19. The induction of ALP was confirmed in donors D23, D24, D26, and D76. E Kinetics of selected genes in response to continuous rhBMP-2 treatment for 28 days. Note that the assay started on day 14 after the cell culture started. n = 3 for each experimental point. The data shown correspond to donor D19. The data were confirmed in donors D23, D24, and D26. F Gene expression profile of selected genes after 28 days of rhBMP-2 treatment. n = 3 for each experimental point. The data shown correspond to donor D19. The data were confirmed in donors D23, D24, and D26. ALPL, alkaline phosphatase biomineralization associated; RUNX2, RUNX Family Transcription Factor 2; OPN, osteopontin; OCN, osteocalcin; ON, osteonectin (stars) indicate statistically significant differences (p < 0.01)