Korean J Transplant.  2022 Nov;36(Supple 1):S218. 10.4285/ATW2022.F-3556.

Role of native liver derived ECM-gel to develop a novel approach for orthotopic hepatocyte transplantation

• Udagawa D
• Yagi H
• Nishi K
• Nagata S
• Hasegawa Y
• Abe Y
• Kitago M
• Kitagawa Y

• KMID: 2535283
• DOI: http://doi.org/10.4285/ATW2022.F-3556

Abstract

Background
In conventional, hepatocytes have been injected via portal vein or directly into liver parenchyma for cell transplantation, however this methodology could not provide efficient cell survival or function in vivo due to their fragile engraftment and dispersion. Recently, novel methodology using hydrogels for hepatocyte transplantation have been introduced to enhance the engraftment and distribution of the cells. However, most commonly used material type I or IV collagen didn’t show improvement of cell survival and function in vivo, since it failed to reproduce the native environment of liver. Therefore, alternative gels are required to develop for improvement of hepatocyte transplantation. This research is aimed to investigate the roles of liver-derived extracellular matrixes gel (L-ECM-gel) in cultivation in vitro and transplantation of human hepatocytes in vivo.
Methods
Solution of L-ECM-gel was prepared by the optimized protocol of decellularization and solubilization of porcine liv-ers. For in vitro assay, human hepatocytes (HepG2 and PXB cells) were dispersed into the hydrogel solution, and immediately gelated in 37 incubator. For in vivo assay, hepatocyte was encapsulated in L-ECM-gel as the graft. The grafts were transplanted between the liver lobes of normal rat. Tacrolimus and Prednisolone were administered as immunosuppressants.
Results
in vitro, the hepatocytes formed cell aggregates by cell-cell interactions during cultivation in hepatocyte medium for 3 days. In quantitative analysis showed production of human albumin (hAlb) in culture supernatant. In vivo, grafts localized at the transplant sites and retained the human hepatocytes. Moreover, hAlb was detected in rat blood, which indicated that hepatocytes demonstrated sufficient functions in the rat body. Furthermore, same results were confirmed in another rat model where Thioacetamide was administered to induce liver fibrosis.
Conclusions
In this study, L-ECM-gel showed the favorable environment for cell engraftment and functionality. Further investigation is required whether L-ECM-gel could help to improve recipients liver fibrosis or damages on hepatocyte transplantation.