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. 2020 Nov 4;30:103–112. doi: 10.1016/j.jare.2020.10.006

Fig. 1.

Fig. 1

KCs-CM improves DP cell inductive phenotype. (a) Amount of active ALP and (b) respective images of ALP-active DP cells showing an improvement of the DP cell inductive-related phenotype after culture with KCs-CM (n = 6). (c) Percentage of DP cells expressing CD184, LRP-4, α-SMA (n = 7) and LEF1 determined by flow cytometry (n = 5, **p < 0.01 vs DMEM). (d) Expression of α-SMA and the V1-isoform of versican in DP cells confirming a decrease after culture with KCs-CM. (e) Representative DAPI-stained nuclei images used to perform image analysis and (f) quantify the number of DP cell aggregates formed after 5 days in culture showing that DP cells treated with KCs-CM have improved capacity to self-aggregate (n = 3) (g) Variation of the area occupied by the DP cells up to 22 h of culture as obtained by the (h) analysis of time-lapse images showing that DP cells cultured in KCs-CM form more compact spheroids and faster than controls (n = 6). (i-k) Amount of growth factors and (l) cytokine secreted by DP cells showing the difference of their secretome with the culture conditions. Data shown are mean ± s.e.m. Differences are indicated by * KCs-CM vs DMEM, # KCs-CM vs Medium CTRL and ● Medium CTRL vs DMEM. *p < 0.05; **p < 0.01; ***p < 0.00; ****p < 0.0001. Scale bars are 50 µm for (b,d), 100 µm for (h) and 200 µm for (e).