A Standardized Murine Model with a Three-Person Workflow for Studying Acute Cellular and Antibody-Mediated Rejection in Xenotransplantation

Despite recent advancements in xenotransplantation, acute rejection remains a major barrier to its clinical application. Acute cellular rejection (ACR) and acute antibody-mediated rejection (AMR) are the primary immune responses leading to early graft failure in xenograft recipients. While large animal models such as non-human primates and genetically modified pigs have provided valuable insights, their use is limited by high costs, ethical constraints, and significant experimental variability. Small-animal models offer a practical, reproducible alternative for mechanistic studies of immune rejection. In this study, two standardized murine xenotransplantation models were developed to specifically mimic ACR and AMR. To ensure reproducibility and minimize operator-related variability, a structured three-person surgical protocol with an assembly-line workflow was implemented, enabling consistent model generation with high efficiency and quality. The resulting models not only replicate the key immunopathological features of clinical xenograft rejection but also offer a robust platform for investigating immune mechanisms and evaluating targeted immunosuppressive strategies. These models will help accelerate the preclinical development of xenotransplantation therapies.