In vivo liver decellularization has become a promising strategy to study in vivo liver engineering. However, long-term survival after in vivo liver decellularization has not yet been achieved due to anatomical and technical challenges. This study aimed at establishing a survival model of in vivo partial liver lobe perfusion-decellularization in rats. We compared three decellularization protocols (1% Triton X100 followed by 1% sodium dodecyl sulfate [SDS], 1% SDS vs. 1% Triton X100, n = 6/group). Using the optimal one as judged by macroscopy, histology and DNA content, we characterized the structural integrity and matrix proteins by using histology, scanning electron microscopy, computed tomography scanning, and immunohistochemistry (IHC). We prevented contamination of the abdominal cavity with the corrosive detergents by using polyvinylidene chloride (PVDC) film + dry gauze in comparison to PVDC film + dry gauze + aspiration tube (n = 6/group). Physiological reperfusion was assessed by histology. Survival rate was determined after a 7-day observation period. Only perfusion with 1% SDS resulted in an acellular scaffold (fully translucent without histologically detectable tissue remnants, DNA concentration is <2% of that in native lobe) with remarkable structural and ultrastructural integrity as well as preservation of main matrix proteins (IHC positive for collagen IV, laminin, and elastin). Contamination of abdominal organs with the potentially toxic SDS solution was achieved by placing a suction tube in addition to the PVDC film + dry gauze and allowed a 7-day survival of all animals without severe postoperative complications. On reperfusion, the liver turned red within seconds without any leakage from the surface of the liver. About 12 h after reperfusion, not only blood cells but also some clots were visible in the portal vein, sinusoidal matrix network, and central vein, suggesting physiological perfusion. In conclusion, our results of this study show the first available data on generation of a survival model of in vivo parenchymal organ decellularization, creating a critical step toward in vivo organ engineering. Impact Statement Recently, in vivo liver decellularization has been considered a promising approach to study in vivo liver repopulation of a scaffold compared with ex vivo liver repopulation. However, long-term survival of in vivo liver decellularization has not yet been achieved. Here, despite anatomical and technical challenges, we successfully created a survival model of in vivo selected liver lobe decellularization in rats, providing a major step toward in vivo organ engineering.
Keywords: extracellular matrix; in vivo liver engineering; in vivo organ engineering; perfusion decellularization in vivo; polyvinylidene chloride film.