Fig. 6

Preeclamptic hCV-MSCs are less capable of supporting cellular network formation of HUVECs. A, B The cellular network formation assay was performed with HUVECs cultured with different media as indicated (control medium, control medium containing 50% MSC normal medium, or containing 50% supernatants from hCV-MSCs of 1st trimester, term control, or term PE placentas). A Representatives of light microscopic images are shown (green: branches; cyan: twigs; yellow: master segments; red surrounded by blue: nodes surrounded by junctions; blue surrounded by red: master junctions). Scale: 200 μm. B Quantification of, the total segment length (B, 1st graph), total master segment length (B, 2nd graph), total number of junctions (B, 3rd graph), and total number of nodes (B, 4th graph) is shown. The results are based on three independent experiments (n = 15 pictures of each condition per group) and presented as bar graphs with mean ± SEM. C Total RNAs were extracted from hCV-MSCs as indicated for evaluating gene levels of VEGF (left) and PLGF (right). The results are from three experiments and presented as relative quantification of gene expression (RQ) with minimum and maximum range. D Secreted VEGF-A (left) and PlGF (right) were measured in supernatants from hCV-MSCs as indicated via ELISA. The results are based on three independent experiments and presented as bar graphs with mean ± SEM. Unpaired Mann-Whitney U test was used for (B), and Student t test was used in C and D. *p < 0.05, **p < 0.01, ***p < 0.001. E Schematic illustration of the proposed working model. PE associated factors like IL6 or TNFα shorten the primary cilium on hCV-MSCs, impairing their own motility, differentiation and signal transduction, and compromising their stimulatory capabilities in terms of cellular network formation of endothelial cells and growth/differentiation of placental organoids. The loss of functional hCV-MSCs with impaired cilia might be connected with progression of preeclampsia