Cultured Hepatocytes Adopt Progenitor Characteristics and Display Bipotent Capacity to Repopulate the Liver

Cell Transplantation

Article

Irmelin Probst¹  Kirsten Unthan-Fechner¹  Sarah Koenig²  Petra Krause² 

[1] Department of Biochemistry I, Centre of Biochemistry and Molecular Cell Biology, University Medical Centre Goettingen, Goettingen, Germany;Department of General and Visceral Surgery, University Medical Centre Goettingen, Goettingen, Germany;

关键词: Liver cell transplantation;    Differentiation;    Retrodifferentiation;    Hepatocyte culture;    Progenitor cells;    Oval cells;   

DOI  :  10.3727/096368913X664856

 received in 2012-10-30, accepted in 2013-02-22,  发布年份 2014

来源: Sage Journals

【 摘 要 】

Clinical studies have proved the therapeutic potential of hepatocyte transplantation as a promising alternative to whole organ liver transplantation in the treatment of hereditary or end-stage liver disease. However, donor shortage seriously restricts cell availability, and the lack of appropriate cell culture protocols for the storage and maintenance of donor cells constitutes a significant obstacle. The aim of this study was to stimulate mature hepatocytes in culture to multiply in vitro and track their fate on transplantation. Rat hepatocytes isolated nonenzymatically were cultured serum free for up to 10 days. They were stimulated into proliferation in the presence of growth factors and conditioned media from nonparenchymal and hepatocyte culture supernatants, as well as 10 mM lithium chloride (LiCl). Cell proliferation was assessed by determining DNA content. Additionally, the extent of cell differentiation was estimated using immunofluorescence staining of hepatic, biliary, progenitor, and mesenchymal markers and gene expression analyses. Transplantation studies were performed on the Fischer CD26-mutant rat following pretreatment with retrorsine and partial hepatectomy. Proliferating hepatocytes increasingly adopted precursor characteristics, expressing progenitor (OV6, CD133), hepatic lineage (CK18), biliary (CD49f, CK7, CK19), and mesenchymal (vimentin) markers. The supplement of LiCl further enhanced the proliferative capacity by 30%. Transplantation studies revealed extensive repopulation by large donor hepatocyte clusters. Furthermore, bile duct-like structures deriving from donor cells proved to be immunoreactive to ductular markers and formed in close proximity to endogenous bile ducts. Mature hepatocytes reveal their potential to “switch” between phenotypes, adopting progenitor characteristics during proliferation in vitro. Following transplantation, these “retrodifferentiated” cells further expanded in vivo, thereby generating bipotentially differentiated progenies (hepatocytes and bile duct-like structures). This apparent plasticity of mature hepatocytes may open new approaches for cell-based strategies to treat liver disease.

【 授权许可】

Unknown   
© 2014 Cognizant Comm. Corp.

【 图 表 】

Figure 4.

Figure 1.