Human embryonic stem cell-derived endothelial cell product injection attenuates cardiac remodeling in myocardial infarction

Figures

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  FIGURE 1. Timeline and design of human embryonic stem cell-derived endothelial cell product (hESC-ECP) differentiation and the 14-day study.

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  FIGURE 2. Representative images of hESC-ECP differentiation and phenotypic characterization. Day 8 morphology and confluency (A) and representative flow cytometric analysis (B,C) of isotype controls and hESC-ECP stained for CD31 (PECAM1) and CD144 (VE-cadherin).

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  FIGURE 3. 1-Day infarct area following hESC-ECP or media injection. Risk area (%) of LV area below the suture (A), infarct area,% (B) within the risk area, and (C) representative sections 1 d post-infarct in CALM, n = 11, and CALC, n = 10 groups. Student’s t-test; data are presented as individual points and as mean ± SEM.

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  FIGURE 4. Echocardiography outcomes in hESC-ECP and media injected mice. Ejection fraction (A) was preserved in CALC mice compared to CALM at 7 d, but not at 14 d post-injection. Global longitudinal strain (B) was increased at 7 d in both CALC and CALM compared with SHAM, but only in CALM mice at 14 d. End-diastolic volume, EDV (C), and end-systolic volume, ESV (D), were preserved at 7 and 14 d in CALC compared with CALM mice. Mitral valve E/A wave ratio (E) and myocardial performance index (F) were not different amongst groups at either time point. ANOVA with Tukey’s post-hoc test. Data are mean ± SEM.

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  FIGURE 5. Immunohistochemical assessment of mature infarct area and myocardial characteristics. Collagen deposition (A,E) and infarct volume (B,F) were significantly reduced in CALC mice compared with CALM. There were no differences in isolectin B4 perfused vascular density (C,G) or myocardial regeneration (D,H) between the groups. Student’s t-test; data are mean ± SEM. White arrows (D) indicate Edu + Troponin T + cells.

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  FIGURE 6. Ligand–receptor interactions between hESC-ECP constituents and resident cardiac populations. (A) UMAP projection of clusters identified from cardiomyocyte-free scRNA-seq data of C57BL/6J hearts taken either prior to CAL (naive), or 1 or 3 days post-CAL. (B) Ligand–receptor interaction (LRI) analysis between day eight hESC-ECP populations and resident cardiac populations at each timepoint. Heatmaps illustrate log10(number of predicted LRI) between each population. Dot plots of LRIs between hESC-ECP populations and (C) myofibroblasts and (D) NK and macrophages.