Phenotypic and in vivo functional characterization of immortalized human fetal liver cells

Abstract

We report the establishment and characterization of immortalized human fetal liver progenitor cells by expression of the Simian virus 40 large T (SV40 LT) antigen. Well-characterized cells at various passages were transplanted into nude mice with acute liver injury and tested for functional capacity. The SV40LT antigen-immortalized fetal liver cells showed a morphology similar to primary cells. Cultured cells demonstrated stable phenotypic expression in various passages, of hepatic markers such as albumin, CK 8, CK18, transcription factors HNF-4α and HNF-1α and CYP3A/7. The cells did not stain for any of the tested cancer-associated markers. Albumin, HNF-4α and CYP3A7 expression was confirmed by reverse transcription polymerase chain reaction (RT-PCR). Flow cytometry showed expression of some progenitor cell markers. In vivo study showed that the cells expressed both fetal and differentiated hepatocytes markers. Our study suggests new approaches to expand hepatic progenitor cells, analyze their fate in animal models aiming at cell therapy of hepatic diseases.

Key Words::

• fetal liver cells
• hepatocytes
• immortalized
• transplantation

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Acknowledgment

This study was financed by the Gothenburg County Council (LUA-ALF), The Lars Erik Gelins foundation and the IngaBritt and Arne Lundbergs foundation to Suchitra Sumitran-Holgersson.

Author contributions

PBP: Participated in performance of the research and data analysis. No conflict of interest. SB: Participated in performance of the research and data analysis. No conflict of interest. MJ: Participated in performance of the research and data analysis. No conflict of interest. MIK: Participated in performance of the research, writing of paper and data analysis. No conflict of interest. MO: Participated in writing of paper. No conflict of interest. SSH: Participated in research design, writing of paper and data analysis.

Declaration of interest: SSH is a shareholder and board member of NovaHep AB, a company developing hepatocyte-like cell lines for diagnostic and therapeutic purposes.