Xeno-Free Integrated Platform for Robust Production of Cardiomyocyte Sheets from hiPSCs

Figure 1

Optimization of hiPSC cardiac differentiation by WNT signaling modulation using different culture systems. (a) Cardiomyocyte differentiation protocol adapted from Lian et al. [29] using enzyme-free passaging and seeding. WNT signaling activation using CHIR was performed for 24 h, while inhibition using IWP4 was performed for 48 h. (b) Effect of CHIR concentration on cardiomyocyte differentiation efficiency in the culture system TeSR/Matrigel by measuring the cardiomyocyte marker cTnT by flow cytometry at day 15. Error bars, SEM: for 3, 4, and 7 μM, for 5 and 8 μM, for 6 μM. value<0.01, value<0.001 relative to 6 μM (ANOVA with Tukey’s test). (c) Screening of CHIR concentration impact on cardiomyocyte differentiation efficiency in the culture system TeSR/Synthemax measured by flow cytometry at day 15. Error bars, SEM: for 3, 4, and 5 μM, for 7 and 8 μM, for 6 μM. value<0.05, value<0.01 relative to 6 μM (ANOVA with Tukey’s test). (d) Cardiomyocyte (CM) output at day 15 per seeded hiPSC for each system using 6 μM of CHIR. Error bars, SEM: for TeSR/Matrigel (M), for TeSR/Synthemax (Syn). value<0.01 (Welch’s -test).

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