Fig. 5

Prolonged cultivation of hiPSC-MPs under non-osteogenic conditions increases conditioned medium functionality. A) DNA content and B) ALP activity quantification after 14 days of continuous supplementation of MSCs (donor F85) with regular CM, prolonged cultivation CM, CM from pre-differentiated hiPSC-MPs or control medium under non-osteogenic and osteogenic conditions. C) Quantification of TGFβ-1 content. Group labels: CTRL—control, CM—conditioned medium, d14-CM—prolonged cultivation conditioned medium, d14OST-CM—osteogenic pre-differentiation conditioned medium, CTRL + O—control with osteogenic supplements, CM + O—conditioned medium with osteogenic supplements, d14-CM + O—prolonged cultivation conditioned medium with osteogenic supplements, d14OST-CM + O—osteogenic pre-differentiation conditioned medium with osteogenic supplements. Data represents a combination of mean ± SD with individual values (n = 4–5 for A, B; n = 3 for C). Statistically-significant differences between the groups (A, B) were evaluated using one-way ANOVA followed by Tukey’s multiple comparisons test and are marked as: *<0,05; ****<0,0001