Korean J Transplant.  2022 Nov;36(Supple 1):S263. 10.4285/ATW2022.F-4041.

Transplantation of chemically induced liver progenitors as a treatment to ameliorate liver fibrosis

• Fukumoto M
• Hidaka M
• Soyama A
• Hara T
• Miyamoto D
• Matsuguma K
• Matsushima H
• Imamura H
• Adachi T
• Eguchi S

• KMID: 2535490
• DOI: http://doi.org/10.4285/ATW2022.F-4041

Abstract

Background
Hepatocyte-based regenerative therapy has been reported as an alternative treatment to liver transplantation for end-stage liver disease. We have been investigating the reprogramming of mature hepatocytes to liver progenitors by chemical stimulation (chemically induced liver progenitor, CLiP) as a possible liver regenerative therapy considering its proliferative capacity and abundant exosomes.
Methods
Experiment 1 was conducted under the following: for making CLiP, hepatocyte isolated from healthy rats were cultured in YAC medium containing Y-27632 (Rho-associated protein kinase inhibitor), A-83-01 (transforming growth factor type I receptor inhibitor), and CHIR99021 (glycogen synthase kinase-3 inhibitor). CLiP was administered to a diet-induced nonalcoholic steatohepatitis (NASH) mouse model (severe combined immunodeficient mice). We evaluated serum laboratory data and the be-havior of rat CLiP. In addition, liver tissue was subjected to Azan staining and smooth muscle actin (SMA) immunostaining after 8 weeks to compare the degree of fibrosis with that of the control group. Experiment 2 was conducted under the following: using surgical specimens, mature hepatocytes were isolated from human cirrhotic liver and cultured in YAC medium. For each degree of fibrosis (F0–F4), immunostaining was performed for liver progenitor cell markers (epithelial cell adhesion molecule, SRY-like HMG box 9, cytokeratin 19). Additionally, the CLiP creation efficiency was measured by flow cytometry using CD133.
Results
The results for experiment 1 are the following: Azan and SMA area-positive rates and serum alanine aminotransferase levels were significantly reduced in the CLiP-treated group (P<0.05). Rat albumin-positive cells were confirmed cell at 8 weeks. The results for experiment 2 are the following: regardless of the degree of fibrosis, cells cultured with YAC medium expressed hepatic progenitor cell markers. Characteristic of CLiP of the cultured cell was determined using CD133, and approximately 60% of the cells were positive.
Conclusions
Transplantation of rat CLiP suppressed the elevation of liver damage markers and dissolved collagen fibers in the liver in a mouse model representing NASH. We succeeded in producing human CLiP from various background livers. CLiP might become an effective cell source for the treatment of liver diseases.