Precision-Cut Liver Slices as a 3D Tissue Model for Assessing Warm Ischemia-Damaged Rat Livers After Normothermic Machine Perfusion

Normothermic machine perfusion (NMP) enables organ assessment and therapeutic intervention prior to transplantation. Evaluating treatment effects conventionally requires whole-organ transplantation, raising ethical and resource concerns. Precision-cut liver slices (PCLS) represent an established ex situ 3D tissue model, but their utility following NMP has not been characterized. We investigated whether PCLS prepared after NMP remain viable and reflect the injury state of the donor organ. Rat livers were allocated to four groups (n = 4/group): native controls, a warm ischemia injury model, and corresponding groups undergoing 6 h of NMP. PCLS were cultured for 48 h and sampled at 2, 24, and 48 h. Injury markers, synthetic function, metabolic activity, inflammatory and oxidative stress gene expression, and histology were assessed. Warm ischemia induction was confirmed by elevated perfusate injury markers during NMP. Alanine aminotransferase remained significantly higher in non-perfused damage-model slices compared with controls after 48 h of culture. Aspartate aminotransferase and lactate dehydrogenase did not differ. Metabolic activity and urea release persisted until 48 h. TUNEL positivity distinguished damage from control slices at 2 h but converged by 48 h. PCLS prepared after NMP remained viable and retained measurable injury- and perfusion-related differences, establishing a novel, 3R-compliant platform for perfusion research.