Establishment of a hepatocyte line from human with SV40LT antigen / 中华传染病杂志

ABSTRACT

Objective To establish a hepatocyte line from human for bioartificial liver support system and hepatocyte transplantation. Methods Hepatocytes isolated from donor liver of a 25 year old brain death male were transfected with pcDNA3.1(-) recombined vector containing the genes encoding Simian Virus 40 large T antigen. Characteristics of the immortalized hepatocyte line were evaluated by morphologic and functional methods. Results One of the hepatocyte clones displayed highly differentiated liver function and immortalized characteristics has been screened after 42 day's selection of 700～300 ?g/ml G418. The immortalized hepatocyte appeared epithelial and displayed morphologic characteristics of liver parenchymal cells. ALB, AST, ALT and LDH secreted from hepatocyte line were detected. Transmission electron microscope examination revealed that immortalized cells contained a round nucleus with nucleoli, moderate numbers of mitochondria, numerous rough endoplasmic reticulums, and some vacuoles. Adjacent cells were joined with junctional complexes. RT PCR and Western blot detection revealed that immortalized hepatocyte could express ALB mRNA and showed immunoreactive of cytochrome P450 2E1. Conclusions The new established hepatocyte line from human retained some specificbiologic functions and morphological cha racteristics of primary cultured hepatocytes.