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Existing iPSC differentiation protocols don\u2019t go beyond producing fetal iCMs. Recently, adult extracellular matrix (ECM) was shown to retain tissue memory and have some success driving tissue-specific differentiation in unspecified cells in various organ systems. Therefore, we focused on investigating the effect of adult human heart-derived extracellular matrix (ECM) on iPSC cardiac differentiation and subsequent maturation. By preconditioning iPSCs with ECM, we tested whether creating cardiac environments around iPSCs would drive iPSCs toward cardiac fate and which ECM components might be involved. We report novel high- and low-abundance proteomes of young, adult, and aged human hearts, with relative abundances to total proteins and each other. We found that adult ECM had extracellular galactin-1, fibronectin, fibrillins, and perlecan (HSPG2) which are implicated in normal heart development. We also showed preconditioning iPSCs with adult cardiac ECM resulted in enhanced cardiac differentiation, yielding iCMs with higher functional maturity, more developed mitochondrial network and coverage, enhanced metabolic maturity, and shift towards more energetic profile. These findings demonstrate the potential use of cardiac ECM in iCM maturation and as a promising strategy for developing iCM-based therapies, disease modeling, and drug screening studies. Upon manipulating ECM, we concluded that the beneficial effects observed were not solely due to the ECM proteins, which might be related to the decorative units attached.<\/jats:p>","DOI":"10.1093\/stmcls\/sxaf005","type":"journal-article","created":{"date-parts":[[2025,1,25]],"date-time":"2025-01-25T14:17:59Z","timestamp":1737814679000},"source":"Crossref","is-referenced-by-count":14,"title":["Adult human heart extracellular matrix improves human iPSC-CM function via mitochondrial and metabolic maturation"],"prefix":"10.1093","volume":"43","author":[{"given":"S","family":"Gulberk Ozcebe","sequence":"first","affiliation":[{"name":"Bioengineering Graduate Program, University of Notre Dame , Notre Dame, 46556 IN ,","place":["United States"]},{"name":"National Institute of Environmental Health Sciences (NIEHS) , Durham, 27709 NC ,","place":["United 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