Figure 2.

PGPC iPSCs Differentiate to Beating Contractile Cardiomyocytes

(A) Differentiation scheme to generate CMs using STEMdiff Cardiomyocyte Differentiation Kit. iPSCs were dissociated to single cells, plated in 12-well plates, and allowed to reach 85%–90% confluency before beginning differentiation.

(B) D16 CMs were dissociated to single cells for reseeding and a proportion was labeled with anti-cTNT-fluorescein isothiocyanate and subjected to flow cytometry (independent experiments ≥3).

(C) Representative images of immunocytochemistry staining of D30 PGPC17 CMs labeled with DAPI, anti-MLC2V (both), and anti-cTNT (left) or anti-α-actinin (right) (independent experiments = 2). Scale bars represent 100 μm. Color channels were independently altered to adjust contrast for publication.

(D) Representative traces of spontaneous Ca²⁺ transients of PGPC CMs at D31 measured by relative fluorescence intensity (independent experiments n ≥ 3; technical replicates per batch ≥2).

(E and F) Plots of (E) beat rate and (F) Ca²⁺ transient amplitudes.

(G) Representative xCELLigence data of D40 PGPC17 CMs showing impedance changes (BAmp: defined as the cell index value between lowest and highest points within a beat waveform) reflecting CM beat waveform and absolute extracellular voltage tracings over a 20-s recording (independent experiments = 3; technical replicates ≥3).