Post-Liver Transplantation Management and Specimen Collection in Daurian Ground Squirrels for Single-Cell RNA Sequencing

Graft functional recovery and graft-host immunocompatibility represent major prognostic concerns following liver transplantation. Critically, conventional rodent models using rats or mice offer limited insight into the mechanisms of hypothermia-induced graft injuries during preservation. This gap underscores the need for alternative models. Since liver grafts from the cold-adaptive Daurian ground squirrel (DGS; Spermophilus dauricus) can sustain prolonged cold preservation and hence support high survivability in post-transplantation DGSs, consequently, this protocol details the essential post-surgical animal care and physiological monitoring required for transplanted DGS subjects. Furthermore, it provides standardized procedures for preparing high-quality liver graft samples for downstream analytical techniques, including single-cell RNA sequencing (scRNA-seq) for transcriptomic profiling and histopathological examination. Successfully generating well-prepared DGS graft specimens is the crucial initial step in applying both traditional and cutting-edge methodologies to study this unconventional yet highly relevant model. This protocol addresses the multi-omic information for DGS, thereby facilitating future research on surgical interventions and disease modeling not only in hibernating rodents but also in other specialized organisms.