A simplified and reproducible ex vivo model of cold and ischemia-reperfusion injury

Abstract

Both cold stress and ischemia-reperfusion injury significantly contribute to poor prognosis after liver transplantation (LT). However, limited animal models incorporating both stimuli hinder the advancement of transplant-related research. Here, a simplified and reproducible isolated perfused liver model is established to simulate the stresses experienced by livers maximally during transplantation. We provide a detailed protocol for a straightforward technique that requires 20-30 min for harvesting, 24-48 h for static cold storage (SCS), and 2 h for normothermic machine perfusion (NMP) to induce LT-like stresses in the liver. Hepatic injury from SCS and NMP (LT-like stresses) is evaluated using three types of parameters. The pH values and hepatic enzyme levels of cold preservation solutions and perfusate serve as dynamic indicators of hepatic injury. Bile production and portal venous resistance directly reflect liver function, whereas pathological analysis visually illustrates the location and extent of injury. This animal model eliminates the influence of hemodynamic and immune factors, yielding highly reproducible results, and is strongly recommended as a standardized animal model for inducing LT-like stresses.

Keywords

Liver transplantation / Animal model / Ischemia-reperfusion injury / Static cold storage (SCS) / Normothermic machine perfusion (NMP) / Ex vivo rat model

Authors’ contributions

Lele Zhang: Writing e review & editing, Writing e original draft. Mingjie Ding: Writing e review & editing. Ying Zhu: Writing e review & editing. Zhiping Yan: Writing e review & editing. Wenzhi Guo: Writing e review & editing, Supervision and Funding acquisition.

Data availability

The data contained in this manuscript or supplementary ma-terial will be made available upon request.

Declaration of competing interest

The authors declare that they have no conflict of interest.

Acknowledgements

This work was supported by the National Natural Science Foundation of China (No. 82400773, 82400745, 82370646), Young Talent Support Program of Henan Association for Science and Technology (No. 2025HYTP083). We acknowledge assistance with the access of analytic instruments from Henan Key Laboratory of Digestive Organ Transplantation and Translational Medical Center of The First Affiliated Hospital of Zhengzhou University. Graphic illustrations were created with BioRender.com.

Appendix A. Supplementary data

Supplementary data to this article can be found online at https://doi.org/10.1016/j.livres.2025.04.005.

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