Figure 1.

Improved structural organization through rod-shaped micropatterning

(A) Schematic overview of the experimental study. On day 35 of differentiation, hiPSC-CMs were seeded onto rod-shaped micropattern or non-patterned slides for 7 to 10 days before molecular (real-time qPCR and immunofluorescence [IF]) and electrophysiological (patch-clamp and transient calcium recordings) phenotyping.

(B) Representative phase-contrast microscopy images of rod-shaped hiPSC-CMs. Images were taken with ×4 (left) and ×10 (right) objectives, scale bar, 100 μm.

(C) Representative IF images of hiPSC-CMs from three different hiPSCs lines (C01, C02, and C03), for alpha-sarcomeric actinin (green) and cardiac troponin T2 (red) under rod-shaped micropatterning (top) vs. non-patterned conditions (down), 7 days after seeding. DAPI (blue) was used to stain nuclei. Scale bar, 50 μm.

(D–H) Violin plots showing quantification data for cell length/width ratio (D), cell circularity index (E), sarcomere organization (F), cell-sarcomere misalignment (G), and sarcomere length (H). Non-patterned conditions, N = 88 for C01, N = 70 for C02, N = 58 for C03; rod-shaped micropattern conditions, N = 47 for C01, N = 58 for C02, N = 47 for C03.

Data are expressed as mean of 3 independent experiments from three different hiPSCs lines. Mann-Whitney: ^∗p < 0.05, ^∗∗p < 0.01 and ^∗∗∗∗p < 0.0001.