| Project/Area Number |
06660414
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Applied veterinary science
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| Research Institution | Nippon Veterinary Animal Science University |
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Principal Investigator |
WASHIZU Tsukimi Nippon Veterinary Animal Science University, Vet.Anim.Sci.Assis.Prof., 獣医畜産学部, 講師 (20191736)
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| Co-Investigator(Kenkyū-buntansha) |
ISHIDA Takuo Nippon Veterinary Animal Science University, Vet.Anim.Sci.Assoc.Prof., 獣医畜産学部, 助教授 (30143506)
ARAI Toshiro Nippon Veterinary Animal Science University, Vet.Anim.Sci.Assis.Prof., 獣医畜産学部, 講師 (70184257)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1995: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1994: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Keywords | primary culture / hepatocytes / cats / glucose metabolism / glucose transport / ヘキソキナーゼ |
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| Research Abstract |
Glucose transport activity and its metabolism of the liver were studied using cultured feline hepatocyes.
Feline hepatocytes were isolated by collagenase perfusion and cultured in Williams medium E with 10% FCS,10^<-7> M insulin, 10^<-7> M dexamethasone, 5000 KIU/I aprotinin in collagen-coated dishes under 37ﾟC in a humid atmosphere of 5% CO_2 in air. The metabolic functions of the cultured feline hepatocytes were evaluated by urea formation, gluconeogenesis and albumin synthesis. The viability of the isolated hepatocvtes was 80.3-87.5% and the yield was 5.1-18.9*10^6 cells/g liver. The hepatocytes became firmly attached to the dishes within 4h after the culture started, spreaded within 24h, formed confluent monolayr within 3 days, and were maintained at least for 7 days. The metabolic functions were well preserved at least for 5 days. Therefore, this primary cultured feline hepatocytes were recognized as a useful in vitro system for several biochemical studies.
Additionaly, we investigated the activities of cytosol enzymes for the glucose metabolism of the feline hepatocytes during culure. The activities of hexokinase, pyruvate kinase, glucose 6-phosphate dehydrogenase, and lactate dehydrogenase were measured as glycolytic enzymes. The activities of these enzymes were well preserved for 7 days.
Also, we studied the sinusoidal lymphocytes isolated by collagenase perfusion. Many of the lymphocytes obtained from both the perfusates and the digested liver were large granular cymphocytes.
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