Transplantation of chemically-induced liver progenitor cells ameliorates hepatic fibrosis in mice with diet-induced nonalcoholic steatohepatitis

Shunsuke Murakami; Akihiko Soyama; Daisuke Miyamoto; Takanobu Hara; Kunihito Matsuguma; Hajime Imamura; Hajime Matsushima; Takayuki Tanaka; Yasuhiro Maruya; Tomohiko Adachi; Satoshi Miuma; Masaaki Hidaka; Kengo Kanetaka; Takahiro Ochiya; Susumu Eguchi

doi:10.1016/j.reth.2022.11.001

Transplantation of chemically-induced liver progenitor cells ameliorates hepatic fibrosis in mice with diet-induced nonalcoholic steatohepatitis

Regen Ther. 2022 Nov 25:21:574-583. doi: 10.1016/j.reth.2022.11.001. eCollection 2022 Dec.

Authors

Affiliations

¹ Department of Surgery, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
² Department of Gastroenterology and Hepatology, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan.
³ Department of Molecular and Cellular Medicine, Tokyo Medical University, Tokyo, Japan.

Abstract

Chemically-induced liver progenitors (CLiPs) have promising applications in liver regenerative medicine. We aimed to clarify the efficacy of CLiPs for ameliorating fibrosis in a diet-induced nonalcoholic steatohepatitis rat model, since nonalcoholic fatty liver disease is currently recognized as the most common form of chronic liver disease in developed countries.

Methods: Primary mature hepatocytes were isolated from 7-week-old male Wistar rats. To establish CLiPs, isolated hepatocytes were cultured in differentiation medium composed of Y-27632, A-83-01, and CHIR99021 (YAC medium). As an animal model that reproduces NASH pathophysiology, 6-week-old severe combined immunodeficient (SCID) mice were carefully selected and prepared and fed with choline-deficient, L-amino acid-defined, high-fat diet (HFD). After 12 weeks' HFD feeding, the mice were assigned to continue HFD with or without the administration of rat CLiPs (HFD + CLiPs and HFD-CLiPs, respectively). Rat CLiPs were administered from the spleen. Hepatic fibrosis was semi-quantitatively evaluated according to histology. Liver parenchyma and blood samples were collected for biochemical analyses.

Results: Rat CLiPs were positive for CK19 and EpCAM were successfully delivered to the liver. At 8 weeks after CLiPs transplantation, the HFD + CLiPs group showed significantly less positive staining than the HFD-CLiPs group. Alanine aminotransferase significantly improved in the HFD + CLiPs group, as demonstrated by Azan staining and αSMA immunostaining. RT qPCR showed that the liver expression of MMP2 and 9 tended to be higher in the HFD + CLiPs group.

Conclusions: The anti-fibrotic effect of CLiPs was demonstrated in the immunodeficient NASH animal model and may have therapeutic applications in humans.

Keywords: Cell transplantation; Chemically induced; Hepatic fibrosis; Hepatocytes; Liver progenitor cells.