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

Figure 2

Replating step increased cell sheet integrity and cardiomyocyte output. (a) Modification of the WNT signaling modulation protocol by adding a replating step at day 6. (b) After initial WNT modulation steps (I.), addition of the replating step increased CM sheet output (IIa.) and for xeno-free systems (e.g., TeSR/Synthemax) reestablished the CM sheet after severe detachment (IIb.) by promoting the proliferation of cardiac progenitor cells (CPCs) (III.). (c) Replating ratios of 1 to 2 and 1 to 3 had no significant impact on differentiation efficiency in the culture system TeSR/Matrigel compared with differentiation without replating (Control). Error bars, SEM: for Control (6 μM) and 1 : 3, for 1 : 2. (d) On the other hand, reestablishing the cell sheet (ratio 1 : 1) in TeSR/Synthemax provided an increase in differentiation efficiency compared with differentiation without replating (Control). Error bars, SEM: for Control (6 μM) and 1 : 2, for 1 : 1. value<0.05 (Welch’s -test). (e) CM output at day 15 per seeded hiPSC increased with replating for the system TeSR/Matrigel. Error bars, SEM: for Control (6 μM) and 1 : 2, for 1 : 3. value<0.05 (Welch’s -test). (f) CM output at day 15 per seeded hiPSC increased significantly with the replating step by reestablishing the cell sheet for the system TeSR/Synthemax. Error bars, SEM: for Control (6 μM), for 1 : 1, for 1 : 2. value<0.05, value<0.01 (Welch’s -test).

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