Fig. 3

Characterization of DP cells co-cultured with EpSlKCs. a Schematic representation of the co-culture established with DP cells and EpSlKCs and the respective monoculture controls. b Representative images of DP cell morphology after staining against the mesenchymal cytoskeleton marker vimentin. c DNA quantification of DP cells along the culture time (n = 5). d Expression of the versican V1 and V2 isoforms by DP cells in the different culture conditions at day 5 of culture. The presence of versican V2-positive cells was observed when DP cells were co-cultured with EpSlKCs (zoomed-in image). Images were counterstained with DAPI. e The inductivity of DP cells was assessed through the quantification of active alkaline phosphatase (ALP, n = 3). f Quantitative flow cytometry data (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; ··p < 0.01 and ····p < 0.0001 vs. day 5; ^#p < 0.05, ^##p < 0.01, and ^####p < 0.0001 vs. day 9. Scale bars are 50 μm