Fig. 3: HiPM-cm promotes human cardiomyocyte proliferation. | Nature Communications

Fig. 3: HiPM-cm promotes human cardiomyocyte proliferation.

From: Medium from human iPSC-derived primitive macrophages promotes adult cardiomyocyte proliferation and cardiac regeneration

Fig. 3

a Schematic diagram depicting the incubation of human cardiomyocytes with fresh culture medium (FM group) or conditioned medium collected from hiPMs (hiPM group). Cartoon elements used in the schematic diagram were created using BioRender. b Daily cell count in human cardiomyocytes treated with FM or hiPM-cm for 4 days. n = 4 per group. c Cardiomyocyte proliferation monitored by IncuCyte instrument at a 12 h interval for 4 days. n = 4 per group. Scale bar, 50 μm. d Immunofluorescence staining of Ki67-positive human cardiomyocytes treated with FM and hiPM-cm in 2D culture system. Human cardiomyocytes were labeled with cTnT (red) and Ki67 (green); nuclei were counterstained with Hoechst (blue). Scale bar, 10 μm. n = 3 biological repeats per group. e Cross-sectional areas of cardiac spheres composed of human cardiomyocytes treated with FM and hiPM-cm at 10 days after the start of 3D culturing. Scale bar, 100 μm. n = 8 spheres for FM group, n = 7 spheres for hiPM-cm group. f Immunofluorescence staining of Ki67-positive human cardiomyocytes treated with FM and hiPM-cm in the 3D culture system. Human cardiomyocytes were labeled with cTnT (red) and Ki67 (green); nuclei were counterstained with Hoechst (blue). Scale bar, 15 μm. n = 3 biological repeats per group. g Immunofluorescence staining of αSMA-positive human cardiomyocytes treated with FM and hiPM-cm. Human cardiomyocytes were labeled with cTnT (red) and αSMA (green); nuclei were counterstained with Hoechst (blue). Scale bar, 10 μm. n = 3 biological repeats per group. h Extracellular acidification rate (ECAR) analysis in human cardiomyocytes treated with FM and hiPM-cm. n = 4 biological repeats per group. i Oxygen consumption rate (OCR) analysis in human cardiomyocytes treated with FM and hiPM-cm. n = 3 biological repeats per group. j ATP levels in human cardiomyocytes treated with FM and hiPM-cm. n = 6 biological repeats per group. k Calcium handling analysis in human cardiomyocytes treated with FM and hiPM-cm. n = 12 cardiomyocytes per group. Quantitative data are presented as the mean ± SEM. Groups were compared using a two-tailed unpaired Student’s t test (b–k). Source data are provided as a Source Data file.

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