Figure 2
From: Cardiac Pacemaker Cells Generate Cardiomyocytes from Fibroblasts in Long-Term Cultures
Characterisation of synchronised and spontaneously beating cell clusters formed during SANC culture. (A) Reverse transcription-polymerase chain reaction (RT-PCR) analysis of cardiac gene expression in cell clusters after culturing SANCs for 3 weeks; Nkx2.5, GATA4, cTnT, desmin, MLC4, HCN4, SERCA2, RYR2 and ANF transcripts, but not those of MLC3, were detected in beating cell clusters during the culture of SANCs (lane ‘CS’). To compare expression levels of cardiac genes in cell clusters, GAPDH and cTnT were used as internal controls in panels a and b, respectively. SA, sinoatrial node tissue; A, atrial cell suspension; V, ventricular cell suspension; CS, cultured SANCs; DF, dermal fibroblasts. Full-length gels from which the images were cropped are given in Supplementary Figs S7–S9. (B) Immunocytochemical detection of cardiac proteins at 2 weeks of culture; cell clusters that had grown around SANCs expressed cTnT (a), desmin (b), KvLQT1 (c), SERCA2 (d), RYR2 (e) and ANF (f). (C) Fine striated sarcomeric patterns of cTnT (green) and actin (red) were observed after 3 weeks of culture; bars, 50 µm (B); 10 µm (C).
