Protocol to generate immunocompetent human heart-macrophage assembloids for the study of immune-cardiac interactions

Summary

Here, we present a protocol to generate fully human pluripotent stem cell (hPSC)-derived human heart-macrophage assembloids. We describe steps for differentiating hPSCs into monocytes, generating cardiac organoids, integrating CD14+ monocytes to become cardiac tissue-resident macrophages, and maturing the assembloids with metabolic cues. Furthermore, we provide a use-case example of disease modeling for the study of atrial arrhythmias. The system provides a reproducible, scalable, and efficient platform for studying immune-cardiac interactions, inflammation, and arrhythmogenesis. For complete details on the use and execution of this protocol, please refer to O'Hern et al.1. Copyright © 2026 The Author(s). Published by Elsevier Inc. All rights reserved.

Authors O'Hern C, Dionise M, Caywood S, Sewavi ML, Basrai H, Muniyandi P, Volmert B, Kostina A, Huang A, Chalfoun N, Aguirre A
Journal STAR protocols
Publication Date 2026 Mar 20;7(1):104411
PubMed 41790550
PubMed Central PMC12992950
DOI 10.1016/j.xpro.2026.104411

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