| Project/Area Number |
06454300
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Pediatrics
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| Research Institution | Tottori University |
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Principal Investigator |
SHIRAKI Kazuo Tottori University, Faculty of Medicine, Professor, 医学部, 教授 (60010229)
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| Co-Investigator(Kenkyū-buntansha) |
NAGATA Ikuo Tottori University, Faculty of Medicine, Research associate, 医学部付属病院, 助手 (50252846)
KONO Yumi Tottori University, Faculty of Medicine, Research associate, 医学部付属病院, 助手 (50243390)
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| Project Period (FY) |
1994 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥5,700,000 (Direct Cost: ¥5,700,000)
Fiscal Year 1996: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1995: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1994: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | THREE-DIMENSIONAL CULTURE / NEONATAL HEPATOCYTE / CHOLESTASIS / 培養肝細胞 / 新生仔 / 新生児 / スフェロイド肝細胞 / 毛細胆管 / 母乳 / 上皮増殖因子 / 肝内胆汁うっ滞 / エストラジオール / EGF |
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| Research Abstract |
We have investigated on the mechanisms of intrahepatic cholestasis to elucidate developmental changes of liver-specific functions in the immature liver using neonatal rat hepatocytes in primary culture. In this study, we developed a spheroidal culture model of neonatal hepatocytes which resembles three-dimensional structure in vivo and examined hepatocellular functions such as protein secretion and formation of bile canaliculi in the model.
Neonatal hepatocytes formed semispheroids after 5 days cultured on dishes coated with polyhydroxyethyl methacrylate. They formed complete spheroids on the dishes coated with BSA.Spheroidal cultured hepatocytes maintained higher levels of albumin secretion than plain cultured hepatocytes after 14 days in culture. Hepatocytes cultured in collage gel sandwich culture system, which provides three-dimensional structures as in vivo, also maintained cuboidal shape of hepatocytes and high levels albumin secretion for 14 days. Bile canaliculilike structure was observed between adjacent hepatocytes by actin staining using rhodamine-phalloidin. Cholestatic agents caused dilatation of these canaliculi.
Thus, three-dimensional culture of neonatal hepatocytes was established. As hepatocytes maintain higher of liver-specific function and morphology, these culture systems should be useful for studying the mechanisms of hepatic cholestasis in the neonatal period.
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