In vivo transplantation of intrahepatic cholangiocyte organoids with decellularized liver-derived hydrogel support hepatic cellular proliferation and differentiation in chronic liver injury

Abstract

Limited replicative potential of primary hepatocytes (Hep) is a major hurdle for obtaining sufficient quantity and quality hepatocytes during cell therapy in patients with liver failure. Intrahepatic cholangiocyte organoids (ICO) derived from intrahepatic bile ducts differentiate into both hepatocytes and cholangiocytes in vitro. Here, we studied in vivo effects of transplanting ICO and Hep in chronic liver injury mice models. Well characterized primary mice ICO and Hep were mixed in decellularized liver matrix hydrogel (DCL) and injected into the subcapsular left lateral liver lobe of CCl4 induced liver injury models whereas mice given DCL alone was the sham group. Two weeks post-transplantation, transplanted liver lobes were collected and studied by histology and RNA sequencing. Transplanted animals did not exhibit any tumors, mortality or morbidity. Mice livers transplanted with ICO had increased cellular proliferation and vascularization as compared to Hep transplanted mice or sham. Collagen deposition in liver was significantly reduced and serum albumin levels was significantly increased in transplanted groups compared to sham. Expression of genes associated with hepatocyte differentiation was highest in Hep transplanted livers among the three groups, but they were also upregulated in ICO transplanted liver in comparison to sham. Our study demonstrates that ICO encapsulated in DCL when transplanted in chronically-injured mice livers engraft well, show hepatocyte differentiation and reduction of fibrosis indicating that hydrogel transplanted cholangiocyte organoids may serve as an efficient cell source and therapy for renewal of hepatocytes, restoration of hepatocyte functions and resolution of liver injury.