| Co-Investigator(Kenkyū-buntansha) |
KOJIMA Hisanori GIFU UNIVERSITY SHOOL OF MEDICINE, DEPARTMENT OF NEUROLOGY AND PSYCHIATRY, CLINICAL RESEARCH FELLOW, 医学部・附属病院, 医員
SHIDOJI Yoshihiro SIEBOLD UNIVER-SITY NAGASAKI, LABORATORY OF CELLLLAR BIOCHEMISTRY.PROFESSOR, 看護栄養学部, 教授 (00111518)
MORIWAKI Hisataka GIFU UNIVERSITY SHOOL OF MEDICINE, FIRST DEPARTMENT OF INTERNAL MEDICINE, PROFESSOR, 医学部, 教授 (50174470)
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| Research Abstract |
For the development of a bioartificial liver (BAL) support device, it is most important to establish highly differentiated liver cells cultured at high density. A hybrid liver support system was developed, and consisted of plasma perfusion through porous hollow fiber modules inoculated with 10 billion porcine hepatocytes entrapped in Engelbreth-Holm-Swarm (EHS) gel. This system was applied to pigs with ischemic liver failure 8 hours after creation of a portocaval shunt and hepatic devascularization. In animals treated with the BAL support system, blood bicarbonate levels were incrcased immediately after treatment, and hemodynamic stability was improved. In control pigs, on the other hand, blood bicarbonate levels and blood pressure remained low. Plasma levels of ammonia and lactate decreased in pigs treated with the BAL device, but not in control animals. We constructed adenovirus vector carring rat HNF-4α cDNA, and transfected the adenovirus vector to hepatoma cells to enforce expression of the exogenous HNF-4α gene. We analyzed expression of HNF-4, HNF-1, and liver specific genes in cells infected by the adenovirus vector expressing HNF-4α by Northem blotting and Western blotting analysis. Adenovirus-mediated HNF-4α gene transfer resulted in the increases of HNF-4, HNF-I, and liver specific genes such as αl-antitrypsin, apolipoproteins, and glutamine synthetase by transformed hepatoma cells. Cells overexpressing HNF-4α removed ammonia from medium supplemented with NH_4Cl to greater extent than cells infected control vector. These results suggest that the use of the BAL support device in combination with a hollow fiber module and hepatocytes entrapped in EHS gel has potential advantages for clinical use in patients with fulminant hepatic failure and that the use of liver cells infected by adenovirus expressing HNF-4a has potential advantages for the development of bioartificial liver.
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