| Project/Area Number |
11557107
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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 | 展開研究 |
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| Research Field |
Cerebral neurosurgery
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| Research Institution | Nagoya City University |
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Principal Investigator |
ASAI Kiyofumi (2000) Department of Bioregulation Research, Nagoya City University Medical School, Associate Professor, 医学部, 助教授 (70212462)
加藤 泰治 (1999) 名古屋市立大学, 医学部, 教授 (60094364)
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| Co-Investigator(Kenkyū-buntansha) |
MIURA Yutaka Department of Bioregulation Research, Nagoya City University Medical School, Assistant Professor, 医学部, 助手 (90285198)
浅井 清文 名古屋市立大学, 医学部, 助教授 (70212462)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥12,800,000 (Direct Cost: ¥12,800,000)
Fiscal Year 2000: ¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 1999: ¥8,800,000 (Direct Cost: ¥8,800,000)
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| Keywords | Blood-Brain Barrier / Endothelial Cell / Astrocyte / BBBモデル / 不死化 / アストロサイト産生因子 |
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| Research Abstract |
(1) Establishment of immortal bovine brain endothelial cell
To establish immortal brain endothelial cell, pSV3-neo which contains SV40 T antigen was introduced to primary cultured bovine brain endothelial cells. After selection by G418, we obtained six clones that exhibited the spindle-shaped appearance of normal brain endothelial cell. One of them, t-BBEC-117, retained the brain endothelial cell phenotype.
(2) Establishment of immortal rat astrocyte
pSV3-neo was introduced to rat primary cultured astrocytes. We obtained four immortal colony, which exhibited glial fibrillary acidic protein. A single clone, ACT-57, exhibited stable growth in a chemically defined medium and a stronger expression of NGF mRNA than that of normal astrocyte.
(3) Identification of astrocytic factor which induce blood-brain barrier phenotype.
Astrocyte in co-culture was found to tighten the intercellular contacts of t-BBEC-117. Among known astrocytic factors, only fibroblast growth factor-basic (bFGF) could mimic the actions of astrocytes. These data suggest that bFGF is one of the most plausible astorcytic factors to induce the blood-brain barrier properties of immortal brain endothelial cells.
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