| Project/Area Number |
15390047
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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 |
Medical pharmacy
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| Research Institution | Shinshu University |
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Principal Investigator |
OHMORI Shigeru Shinshu University, Shinshu University Hospital, Division of Pharmacy, Professor, 医学部附属病院, 教授 (70169069)
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| Co-Investigator(Kenkyū-buntansha) |
MATSUNAGA Tamihide Shinshu University, Shinshu University Hospital, Division of Pharmacy, Associate Professor, 医学部附属病院, 助教授 (40209581)
SASAKI Katsunori Shinshu University, Shinshu University School of Medicine, Department of Anatomy and Organ Technology, 医学部, 教授 (30170666)
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| Project Period (FY) |
2003 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥14,800,000 (Direct Cost: ¥14,800,000)
Fiscal Year 2005: ¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 2004: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 2003: ¥9,400,000 (Direct Cost: ¥9,400,000)
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| Keywords | mouse / monkey / embryonic stem cell / hepatocyte / differentiate / hepatoblast / cyochrome P450 / CYP3A / CYP3A8 / CYP3A66 / HNF-3β / 胚様体 / 薬物トランスポーター / Cyplal / Cyp3all / Cyp7al / Cyp7bl |
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| Research Abstract |
To differentiate mouse embryonic stem (ES) cells to hepatocytes, embryoid bodies were cultured for 36 days. Hepatocyte differentiation markers were expressed in the cells differentiated from ES cells. The expression pattern of cytochrome P450 (CYP) genes in the cells could be grouped into four different categories, such as highly expressed at early to mid stage, only expressed at late stage ; expressed throughout at all stages and not detectable at any stages. Aryl hydrocarbon receptor, aryl hydrocarbon receptor nuclear translocator and glucocorticoid receptor were detected in the cells cultured throughout the time course. Pregnane X receptor was detected only the cells cultured for more than 24 days. The alpha-fetoprotein and albumin were co-expressed with CYP3A2 and CYP1A1, respectively. Monkey ES cells were also differentiated into hepatocytes in vitro. The cells expressed alpha-fetoprotein, albumin, CYP3A8, CYP3A66 and CYP7A1. We have isolated a cDNA clone from the differentiating monkey ES cells and identified a novel CYP isoform, named CYP3A7m. Deduced amino acid sequence of CYP3A7m shows 89, 81, 91 and 72% homology to CYP3A4, CYP3A5, CYP3A7 and CYP3A43, respectively. These results suggested that CYP and transcription factor genes were expressed in hepatocytes differentiated from ES cells and those expression levels and patterns were considerably changed in individual enzymes during the course of the differentiation.
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