| Project/Area Number |
13558104
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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 |
Biomedical engineering/Biological material science
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| Research Institution | The University of Tokyo |
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Principal Investigator |
IMAMURA Yasutada The University of Tokyo, Graduate School of Arts and Sciences, Assistant Professor, 大学院・総合文化研究科, 助手 (40201339)
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| Co-Investigator(Kenkyū-buntansha) |
HAYASHI Yoshihiko Teikyo Heisei University, Faculty of Information, Professor, 情報学部, 教授 (60090528)
大塚 龍郎 新田ゼラチン株式会社, 生物科学研究所, 所長
水野 一乘 東京大学, 大学院・総合文化研究科, 助手 (70251337)
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| Project Period (FY) |
2001 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥11,600,000 (Direct Cost: ¥11,600,000)
Fiscal Year 2003: ¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2002: ¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2001: ¥4,300,000 (Direct Cost: ¥4,300,000)
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| Keywords | type IV collagen / type IV collagen gel / smooth muscle cell / mesangium cell / collagen / gel / basement membrane / type V collagen / メッシュワーク / コラーゲン会合体 / レンズカプセル / 線維芽細胞 / コラーゲンゲル / 多細胞メッシュワーク |
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| Research Abstract |
This study aimed to explore and establish utilization of type IV collagen as culture substrate and culture method using the collagen. Transfer of some techniques related to manufacturing of the culture substrates was originally intended. Banned distribution of bovine derived material in Japan after first finding of a cattle affected with BSE made us to change our study plan. Since it was impossible to get the bovine sources, especially eye-related tissues such as lens capsules in Japan which is the best starting material for type IV collagen preparations, import of those from BSE-free country was successfully tried. But increased cost for the preparation resulted in small scale preparation only for laboratory use. New finding are summarized as follows :
1.Improved recovery of type IV collagen from bovine lens capsule
We obtained more efficient method with roughly twice amount to recover type IV collagen capable of forming gel. Type IV collagen in the preparation by the new method was par
… More
tially modified with some amino acid residues but still retained gel forming property on which human smooth muscle cells and human mesangium cells formed meshwork like cellular network.
2.Trials of new cell culture methods using type IV collagen
a.A hat shape gel
Type IV collagen gel formed on a circle cover glass for cell culture support were getting dry from its periphery and remained in the center part of the circle. The shape of the substrate looks like a hat which is put on man's head and is called a hat shape gel. Same type IV collagens form two types of molecular aggregates between periphery and center. Smooth muscle cells cultured on the hat shape gel showed remarkable differences between both parts of the culture glass. The cells in the center part formed meshwork such as cells cultured on type IV collagen gel but those in periphery part did not form meshwork but grew well. The result demonstrated that physical property of the type IV collagen aggregates affects cellular response.
b.Utilisation of type V collagen as cell culture substrates
Type V collagen distributes relatively close to basement membrane. Effect of type V collagen aggregates as culture substrates was examined and compared that of type IV collagen aggregates. Type V collagen aggregates affects unique effect on cellular response, such as shape and proliferation. Less
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