Fig. 4. Optimization of human FC yield.

(A) Effect of culture substrate on FC yield. Data in the time-course plot are shown as means ± SEM. The P value was calculated using analysis of variance (ANOVA) (n = 4 to 12 per condition). The phase-contrast images show FC morphology at 14 days. Scale bar, 200 μm. (B) Effect of the PBMC isolation method [Vacutainer CPT (vac) versus ACK lysis (lysis) of red blood cells] on FC yield. The P value was calculated using Student’s t test (n = 6 per condition). The phase-contrast images show FC morphology at 14 days. Scale bar, 25 μm. (C) Venn diagram summarizing proteome coverage in FCs differentiated following vac or lysis (n = 3 per condition). (D) Scatterplot matrix summarizing correlation analysis of log₂ LFQ intensities (relative protein abundances) in FCs differentiated following vac or lysis (n = 3 per condition). Correlation coefficients corresponding to each scatterplot (calculated using Pearson’s r) are represented by the heatmap overlay. (E) Hierarchical clustering analysis of log₂ LFQ intensities in FCs differentiated following vac or lysis (n = 3 per condition). (F) Volcano plot summarizing differential protein abundance in FCs differentiated following vac or lysis (n = 3 per condition). Gray curves denote cutoff criteria, generated in Perseus; P values were calculated using Student’s t test.