Fig. 4

Preeclamptic hCV-MSCs differentiate poorly. A–C hCV-MSCs from 1st trimester, term control, and term PE placentas were subjected to osteogenic differentiation medium for 21 days. The percentage of differentiated osteogenic cells was evaluated by Alizarin Red S staining. A Representative images for Alizarin Red S staining are shown (arrowheads depict calcification). Scale: 20 μm. B The quantification of Alizarin Red S positive cells in percentage is shown as bar graph with mean ± SEM (B, 1st graph) as well as the mean gray value (B, 2nd graph) (n = 15, pooled from three independent experiments with three individual hCV-MSCs). C Expression levels of three osteogenic differentiation related genes PTCH1 (left), OPN (middle), and KLF4 (right) in control and hCV-MSCs upon differentiation. The results are from three independent experiments and presented as RQ with minimum and maximum range. D, E hCV-MSCs from 1st trimester, term control, and term PE placentas were subjected to chondrogenic differentiation medium for 21 days. The percentage of differentiated chondrogenic-like cells was evaluated by quantifying the Alcian Blue positive cells. D Representatives of chondrogenic-like cells stained by Alcian Blue are shown (blue: aggrecans). Scale: 50 μm. E Quantification of the mean gray value in percent (left) and the percentage of chondrogenic-like cells (right) are illustrated as bar graph showing mean ± SEM (n = 15, pooled from three independent experiments with three distinct hCV-MSCs for each group). Student’s t test was used in C. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. Unpaired Mann-Whitney U test was used in B and E. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001