Human cell-based models of cardiac ischaemia-reperfusion injury: a scoping review of experimental procedures

Cardiac ischaemia-reperfusion injury (IRI) is a major contributor to myocardial damage following coronary diseases and cardiac transplantation. Despite extensive preclinical research demonstrating cardioprotective strategies in animal models, successful translation into human therapies has been limited. While animal models capture systemic in vivo complexity, they do not fully replicate human cardiac physiology, and the intertwined nature of signalling pathways in vivo makes mechanistic dissection challenging. Human cell-based in vitro models offer a valuable complementary way to study cardiac ischaemia-reperfusion injury. They allow researchers to tightly control experimental conditions, explore underlying mechanisms in detail, and test potential therapeutic strategies. Despite these advantages, there is substantial heterogeneity in how cardiac IRI is modelled in vitro. Studies differ widely in the cell types used, the methods and durations of ischaemia and reperfusion, oxygen and nutrient conditions, and the outcomes assessed. This lack of consistency makes results harder to interpret, reduces reproducibility, and limits meaningful comparisons between studies. This systematic scoping review focuses on human cell-based in vitro models of cardiac IRI. It aims to comprehensively map the experimental approaches used to simulate ischaemia and reperfusion in human cardiac cells, including human induced pluripotent stem cell-derived cardiomyocytes, primary cardiac cells, and advanced 3D models. The review will synthesise current evidence on commonly employed ischaemia induction methods such as oxygen-glucose deprivation, ischaemic mimetic buffers, and hypoxic chamber exposure as well as reperfusion strategies involving reoxygenation and nutrient restoration. Key methodological features will be systematically examined, including ischaemia and reperfusion durations, oxygen levels, nutrient conditions, and outcome assessment techniques. The review will compare practices across studies, highlighting variability in endpoints such as cell viability, oxidative stress, mitochondrial function, and contractility. It will also evaluate emerging methods, including the use of 3D and multicellular models and the potential for personalised approaches using hiPSC-derived cardiac cell populations. Expected outcomes include a comprehensive mapping of existing methodologies, identification of common practices and inconsistencies, and recognition of critical gaps in current experimental designs. This synthesis will inform researchers about the strengths and limitations of current human cell-based cardiac IRI models and will support efforts toward methodological standardisation. Ultimately, this work aims to enhance reproducibility, improve translational relevance, and facilitate the development of more robust and personalised therapeutic strategies for reducing cardiac IRI.