Production and characterization of immortalized rat hepatocytes secreting hepatocyte growth factor/scatter factor.

Abstract

BACKGROUND/AIMS: Hepatocyte transplantation is a recently attractive field in the treatment of liver failure and enzyme-deficient diseases. However, procurement of sufficient quantities of hepatocytes is almost impossible. We attempted to create a hepatocyte cell line that could be used for hepatocyte transplantation.

METHODOLOGY: L2A2 is a conditionally immortalized rat hepatocyte cell line produced by transfection of temperature-sensitive simian virus T antigen to the hepatocytes in the Lewis rat. Hepatocyte Growth Factor/Scatter Factor (HGF/SF)-secreting L2A2 cells, designated as SF-21, was produced by transfecting human HGF/SF cDNA into L2A2 cells.

RESULTS: This cell line was able to produce HGF/SF at the rate of 5-10 ng/10(6) cells/24 hrs, and the recombinant HGF/SF was of the expected size and was functionally active in that it could scatter Madin-Darby canine kidney cells. The SF-21 cells grew faster than its parental cell clone, and survived and proliferated at 37 degrees C in vitro. Also, the SF-21 cells were able to survive and proliferate when transplanted into the spleen of syngeneic rat, and expressed glucose-6-phosphatase.

CONCLUSIONS: These HGF/SF-secreting hepatocytes can be used as a model system to test a feasibility of using genetically engineered hepatocyte cell line for hepatocyte transplantation in the rat.