Figure 4. ECM requirements for matrix sandwich protocol-based cardiac differentiation and robust application to multiple hPSC lines.
(A) Effect of single or double Matrigel overlays on cardiac differentiation using the protocol defined in Figure 3(A) with Activin A (100 ng/ml), BMP4 (10 ng/ml) and bFGF (5 ng/ml). Cardiogenesis efficiency was measured by flow cytometry for cTnT+ CMs at 15 days differentiation of the iPSC line DF19-9-11T. Control is the cell culture without Matrigel overlay. Error bars represent SD, N=9. (B) Cardiac differentiation of multiple hPSC lines assessed by flow cytometry for cTnT at 15 days differentiaton using the matrix sandwich protocol. Error bars represent SD, N=15 for IMR90 C4; N=15 for DF6-9-9T; N=13 for DF19-9-7T; N=21 for DF19-9-11T; N=24 for H1; N=16 for H9. Data were compared using one-way ANOVA with * indicating significantly different, P < 0.05.