Figure 2
Differences in cardiac differentiation capability among hiPSC lines. (a) Schematic of the culturing process for cardiac differentiation in EB suspension cultures. (b) Comparison of the cardiac differentiation ability of hiPSC lines using flow cytometric analysis at 17 d post-induction of cardiac differentiation with different concentrations of Activin A; 6 ng/mL and 12 ng/mL. Data are expressed as mean ± SEM (n = 3). **p < 0.01 vs. R-2A, ANOVA and Dunnett’s test. (c) Proportion of rhythmic and synchronous beating EBs at 8 d and 17 d post-differentiation with different concentrations of Activin A; 6 ng/mL and 12 ng/mL. Data are expressed as mean ± SEM (n = 3). **p < 0.01 vs. R-2A, ANOVA and Dunnett’s test. (d) Immunofluorescence of hiPSC-derived cardiomyocytes for cardiac-specific markers. Micrographs show cTnT (green), Nkx2.5 (red), and Hoechst (blue) staining. Upper panels show low magnification and lower panels high magnification. Scale bars in upper panels = 300 μm, in lower panels = 100 μm. (e) Expression of an undifferentiated hiPSC marker gene (LIN28) among hiPSC lines at 17 d post-cardiac differentiation. Data are expressed as mean ± SEM (n = 6). *p < 0.05, t-test. All mRNA values are shown as fold change relative to the expression of R-2A in Low differentiation group. (f) Heat map of cardiomyocyte-related genes and maturation-related genes among six hiPSC lines. (g) Principal component analysis of cardiac differentiation ability among six hiPSC lines at 17 d post-cardiac differentiation with different concentrations of Activin A; 6 ng/mL and 12 ng/mL (FC1, first principal component scores).
