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Figure 4 | BMC Developmental Biology

Figure 4

From: Human Dental pulp stem cells (hDPSCs): isolation, enrichment and comparative differentiation of two sub-populations

Figure 4

Evaluation of osteogenic differentiation of STRO-1 + /c-Kit + /CD34 − hDPSCs and STRO-1 + /c-Kit + /CD34 + hDPSCs. A: Confocal analysis of osteogenic differentiation of STRO-1+/c-Kit+/CD34− hDPSCs and STRO-1+/c-Kit+/CD34+ hDPSCs. Double immunofluorescence confocal images showing signals from anti-OPN (red), anti-OCN (green) and DAPI (blue), and from anti-OCN (green) and anti-Osx (red), respectively. Bar: 10 μm. B: Alizarin Red staining on STRO-1+/c-Kit+/CD34− hDPSCs and STRO-1+/c-Kit+/CD34+ hDPSCs. Densitometric analysis show the deposition of extracellular mineralized matrix by both the subpopulations after the osteogenic induction. Values are mean ± SD of gray levels (0–255 scale). STRO-1+/c-Kit+/CD34− hDPSCs n = 3; STRO-1+/c-Kit+/CD34+ hDPSCs n = 3; *** p<0,001 STRO-1+/c-Kit+/CD34− hDPSCs and STRO-1+/c-Kit+/CD34+ hDPSCs vs undifferentiated hDPSCs, ANOVA test followed by Newman-Keuls post-hoc test. C: Western blot (WB) analysis of Coll-I and OCN expression in whole cell lysates of differentiated STRO-1+/c-Kit+/CD34− hDPSCs and STRO-1+/c-Kit+/CD34+ hDPSCs. Whole cell lysates were collected from three plates of hDPSCs for each experimental group. Fibroblasts were used as positive controls for Coll-I expression and as negative controls for osteocalcin expression, respectively. Actin bands demonstrate that an equal amount of protein was loaded in each line. Densitometric analysis of the bands corresponding to Coll-I and OCN is shown below western blot images. Values are mean ± SD of gray levels (0–255 scale); ***p<0,001 diff STRO-1+/c-Kit+/CD34− hDPSCs vs undiff STRO-1+/c-Kit+/CD34−, §§§ p<0,001 diff STRO-1+/c-Kit+/CD34+ hDPSCs vs undiff STRO-1+/c-Kit+/CD34+ hDPSCs, ANOVA test followed by Newman-Keuls post-hoc test.

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