| Co-Investigator(Kenkyū-buntansha) |
KAKISAKA Akitoshi Asahikawa Medical College, Assistant Prof., 医学部, 助手 (60194677)
YAMAMOTO Tetsu Asahikawa Medical College, Assistant Prof., 医学部, 助手 (50125415)
加藤 一哉 旭川医科大学, 医学部, 助手 (70175280)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1989: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1988: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Research Abstract |
We have developed a prototype of hybrid artificial liver support system using isolated hepatocytes and have reported previously. In those systems, the viability of hepatocytes could not be maintained over ten hours because of a floating system. The purpose of this study is to evaluate the usefulness of hepatocyte culture substrata to maintain the viability and functions of hepatocytes for longer time developing a hybrid artificial liver.
Male Wister rats weighing about 2OOg were used to obtain isolated hepatocytes by collagenase digestive method. As natural substrata, collagen, fibronectin, vitronectin and biomatrix were examined. Biomatrix was prepared from dog liver by the method of Rojkind. As synthetic substrata, lactose-carrying polystyrene (PVLA) and microcarrier Cytodex 3 were used, and entrapped hepatocytes within carcium arginate gel was also examined.
Cell attachment of isolated hepatocytes and their behavior were observed microscopically on the dishes coated with collagen, fibronectin, vitronectin or biomatrix, PVLA coated dish and Cytodex 3 were also investigated for the hepaticytes attachment. Metabolic functions were examined in dish culture and in spinner flask incubations.
Hepatocytes on the dish coated with substrata such as collagen, fibronectin, vitronectin attached and spread well 24 hours after seeding, but hepatocytes did not spread on biomatrix. On the dish with PVLA, hepatocytes attached well but not spread, and the hepatocytes also attached on Cytodex 3. In dish culture test and spinner flask test, remarkable decrease of NH_3 concentrations with a little increase of BUN and glucose concentrations were observed in each materials.
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