Fig. 4

Growth curves of isolated SC/MPC populations. a Representative original growth curves of cells from mixed P40/70, SP40/50, and SP50/70 started at day 8 of culture. The impedance-based xCELLigence system was used for real-time monitoring of adhesion and proliferation of SC populations seeded at densities of 5000 cells per well (gray) or 40,000 cells per well (black), respectively. In adhesion assays, growth curves were analyzed regarding the slope during the adhesion phase (A) and the CI decrease after medium change. In proliferation assays, growth curves were analyzed with regard to the doubling time (DT) during the logarithmic growth phase and the time for CI to reach half of its maximum value (50% CI, marked with *). Curves were obtained from quadruplicates measured in parallel for each condition, and representative results are shown as means ± SD. b Box-Whisker plots are shown for selected growth parameters and include all experiments performed (n = 5–12 animals). Whiskers are presented with the maximum 1.5 of the interquartile range (Q₁ – Q₃), and the resulting outliers are included as circles. Statistical analysis of differences in growth parameters was performed by an ANOVA (adhesion slope, 50% of CI_max) or Kruskal-Wallis One-Way ANOVA on Ranks (CI decrease after medium change) followed by a pairwise comparison test, and significant differences are marked with asterisks (*p ≤ 0.05, **p ≤ 0.01, ***p < 0.001). All populations showed characteristic kinetic profiles, but measurable differences occurred between them. The slope during cell adhesion is significantly steeper in mixed P40/70 and SP40/50 in comparison with SP50/70. The CI of mixed P40/70 and SP40/50 is influenced negatively by medium change, but significantly less dramatically in SP50/70. The time needed to reach 50% of the maximum CI (50% of CI_max) value was different among the populations, whereas SP50/70 showed significantly higher values than mixed P40/70 and SP40/50