5.4. Effect of CZE on Cell Growth and Cardiomyocytes Differentiation

To investigate the effect of CZE on cell growth and CMs differentiation, differentiation was performed using the mass culture protocol as previously described [16]. Briefly, embryoid bodies (EBs) were first formed in nonadherent plates by culturing 1 × 10⁶ iPS cells in 14 mL Iscove’s Modified Dulbecco’s Medium (IMDM) supplemented with 20% FCS (Invitrogen) in the absence (untreated) or presence of CZE (CZE-treated) at different concentrations as stated above. Afterwards, untreated and CZE-treated EBs were diluted to a density of about 1000 EBs/12 mL in differentiation medium. Media and CZE were changed every second or third day throughout the differentiation period. EB formation was assessed by general morphology observation and analysis of EB diameters and beating activities (Figure 5A,B).