| Project/Area Number |
18591514
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Digestive surgery
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| Research Institution | Kyoto University |
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Principal Investigator |
FUKUCHI Yoshie Kyoto University, Kyoto University Graduate School of Medicine, Technician (70402906)
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| Co-Investigator(Kenkyū-buntansha) |
IKAI Iwao Kyoto University, Graduate School of Medicine, Associate Professor (60263084)
YASUCHIKA Kentaro Kyoto University, Graduate School of Medicine, Insutuctor (00378895)
HATANO Etsuro Kyoto University, Graduate School of Medicine, Insutuctor (80359801)
待本 貴文 京都大学, 医学研究科, 医員 (70437234)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,650,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥450,000)
Fiscal Year 2007: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2006: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | mesenchymal cells / hepatic stem / progenitor cells / undifferentiated state |
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| Research Abstract |
(1) Establishment of in vitro differentiation of hepatic stem/ progenitor cells with Thy1 (+) gp38 (+)-mesenchymal cells
According to the findings of the upregulation of glycogen deposit, gene expression of mature hepatocyte, and also morphological features of mature hapatocyte determined in the haptic stem/progenitor cells (HPCs) isolated from mouse fetal liver tissue, we identified the in vitro differentiation of HPCs after the co-culture with Thy1 (+) gp38 (+)-mesenchymal cells which were also resided in the fetal liver. This effect was not identified when HPCs were co-cultured with the conditioned medium of Thy1 (+) gp38 (+)-mesenchymal cells. We also performed the comprehensive gene expression analysis corresponding to the maturation of HPCs using microarray analysis. Furthermore, we established the cell lines of Thy1 (+) gp38 (+)-mesenchymal cells with gene transfection of SV40 large T antigen in order to execute stable analysis for the mechanism of in vitro maturation of HPCs.
(2)
… More
Establishment of the maintenance for HPCs with undifferentiated state
1) Culture condition
According to the findings of no expression of mature hepatocyte markers, maintenance of co-expression of hepatocyte and cholangiocyte markers in the HPCs co-cultured with Thy1 (+) gp38 (-)-mesenchymal cells, we identified the maintenance of HPCs with undifferentiated state. Fhrthermore, Brd-U incorporation rate was upregulated in the HPCs when they co-cultured with Thy1 (+) gp38 (-)-mesenchymal cells. This finding suggested that the highly proliferative potential of HPCs was maintained by Thy1 (+) gp38 (-)-mesenchymal cells.
2) Functional analysis of Thy1 (+) gp38 (-)-mesenchymal cells
The in vitro differentiation of HPCs co-cultured with Thy1 (+) gp38 (+)-mesenchymal cells was inhibited when they were cultured in the conditioned medium of Thy1 (+) gp38 (-)-mesenchymal cells. This finding revealed the fact that some liquid factors secreted from Thy1 (+) gp38 (-)-mesenchymal cells would have subtstantial role for the maintenance of HPCs with undifferentiated state. Less
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