Additional file 3 of Human placental mesenchymal stromal cells are ciliated and their ciliation is compromised in preeclampsia

Additional file 3: Figure S3. hCV-MSCs from term PE placentas reduced their ability to support growth of placental organoids and migration of EVT cells. (A) Placental organoids were formed for 72 h by using hTSCCT cells and treated then for up to 7 days with 30% supernatant from 1st trimester, term control or term PE hCV-MSCs. The organoids were stained against β-hCG (red), pHH3 (green) and DNA (DAPI, blue), and their diameters were microscopically evaluated. The results are presented as scatter graphs showing the mean ± SEM (n = 7-10 organoids, from three different hCV-MSCs supernatants for each group). (B-F) Illustration of cell co-culture experiment. HIPEC/HTR cells (blue) were seeded into Ibidi chambers and surrounded by indicated hCV-MSCs from 1st trimester, term control and term PE placentas (red). After 8 h the chambers were removed, and the medium changed to HTR or HIPEC medium. Images were taken at indicated time points (0, 4, 8 h) to document the migration font. (B and D) Quantification of the open area between both migration fronts at various time points, for HTR (B) and HIPEC cells (D). The cell-free area at 0 h was assigned as 100%. The results from three independent experiments are presented as mean ± SEM. Unpaired Mann–Whitney U-test was used. *p < 0.05, **p < 0.01. (C and E) Representatives of the migration front are shown. White dashed line depicts the free area of the migration front. Scale: 200 μm. (G) Total RNAs were extracted from hCV-MSCs for analyzing gene levels of MMP2 (G, left graph) and MMP9 (G, right graph). The data are based on three independent experiments and presented as RQ with minimum and maximum range. RQ: relative quantification of the gene expression. Student’s t-test was used. *p < 0.05