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The LIM-homeodomain transcription factor ISL1 marks multipotent cardiac progenitors that give rise to cardiac muscle, endothelium, and smooth muscle cells. ISL1⁺ progenitors can be derived from human pluripotent stem cells, but the inability to efficiently isolate pure populations has limited their characterization. Using a genetic selection strategy, we were able to highly enrich ISL1⁺ cells derived from human embryonic stem cells. Comparative quantitative proteomic analysis of enriched ISL1⁺ cells identified ALCAM (CD166) as a surface marker that enabled the isolation of ISL1⁺ progenitor cells. ALCAM⁺/ISL1⁺ progenitors are multipotent and differentiate into cardiomyocytes, endothelial cells, and smooth muscle cells. Transplantation of ALCAM⁺ progenitors enhances tissue recovery, restores cardiac function, and improves angiogenesis through activation of AKT-MAPK signaling in a rat model of myocardial …