| Project/Area Number |
12555060
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 展開研究 |
|---|
| Research Field |
Thermal engineering
|
|---|
| Research Institution | Keio University |
|---|
Principal Investigator |
TANISHITA Kazuo Keio University, Faculty of Science and Technology, Professor, 理工学部, 教授 (10101776)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
IKEDA Mariko Keio University, Professor Emeritus, 名誉教授 (00051368)
NAGASAKA Yuji Keio University, Faculty of Science and Technology, Professor, 理工学部, 教授 (40129573)
OKA Kotaro Keio University, Faculty of Science and Technology, Associate Professor, 理工学部, 助教授 (10276412)
MITAKA Toshihiro Sapporo Medical University, School of Medicine, Associate Professor, がん研究所, 助教授 (50231618)
|
|---|
| Project Period (FY) |
2000 – 2001
|
| Project Status |
Completed (Fiscal Year 2001)
|
| Budget Amount *help |
¥13,500,000 (Direct Cost: ¥13,500,000)
Fiscal Year 2001: ¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2000: ¥10,200,000 (Direct Cost: ¥10,200,000)
|
|---|
| Keywords | self-organization / tissue engineering / cell culture / biomechanics / bile canalicular formation / bio-artificial liver / 肝細胞 / 細胞コロニー / 幹細胞 / 分化誘導 |
|---|
| Research Abstract |
The morphogenesis and movement of bile canaliculi (BC) are not well understood. This is because culture of hepatocytes that maintain polarity of cell membranes and possess highly differentiated functions has never been successful. We found that small hepatocytes (SHs), which are known to be hepatic progenitor cells, could proliferate and differentiate into mature hepatocytes and that BC-like structures developed between rising/piled-up cells. In the present experiment we examined how SHs could form BC-like structures in the process of their maturation, whether the structures could be functionally active as BC, how BC-like structures contract to expel substances, and whether A23187 and Endothelin-1 caused BC contractions or not.
The results of immunocytochemistry, immunoblots, and immunoelectron micrographs revealed that BC proteins localized in the intercellular space that coincided with BC-like structures formed between rising/piled-up cells. Fluorescein was secreted into BC and accumulated without leakage. Time-lapse microscopy showed that continuous contraction and dilatation occurred in the BC-like structures, and fluorescent dye was secreted into the reformed BC and unidirectionally carried through the structures. We also found out BC contractions occurred in a coordinated manner, which at least a few cells length of the BC-like structures contract synchronously. It is also found out that both A23187 and Endothelin-1 caused BC contractions. The functional abilities of reformed BC may be equivalent to those of in vivo BC.
|
