| Project/Area Number |
13671268
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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 |
General surgery
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| Research Institution | Fukuoka University |
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Principal Investigator |
ONO Junko Fukuoka University, School of Medicine, Professor, 医学部, 教授 (40108692)
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| Co-Investigator(Kenkyū-buntansha) |
KATSUTA Hitoshi Fukuoka University, School of Medicine, Research Assistant, 医学部, 助手 (50333240)
ANZAI Keizo Fukuoka Univ. Hospital, Lecturer, 講師 (60258556)
YASUNAMI Yohichi Fukuoka University, School of Medicine, Assistant Professor, 医学部, 助教授 (00166521)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2002: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2001: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | Islet hormone-producing cells / Gene expression / Differentiation / cre-loxP system / インスリン / グルカゴン |
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| Research Abstract |
To facilitate the introduction of new strategies, such as transplantation of islet cells, gene therapy and bioartificial pancreas, to the treatment of insulin-dependent diabetes mellitus, the establishment of methods for inducing proliferation and differentiation to islet or islet progenitor cells is an urgent project. Therefore, we tried to clarify the precise processes of differentiation of each type of islet hormone-producing cells by using a cre-loxP system to select cells ideal for transplantation.
At first, we focused on insulin-producing beta cells and constructed an expression vector for cre gene with an insulin-II promoter, and successfully obtained insulin-II promoter-cre transgenic mice. The selective expression of cre gene in insulin-producing cells was confirmed by RT-PCR and immunohistochemistry with an anti-insulin antibody. The second transgenic mice were established, in which the expression of EGFP was restricted only to the cells coexpressed EGFP with a cre recombinase. From these two strains, the double transgenic mice were obtained. Recombination of loxP sites was occurred and the transcription termination cassette was cut down in pancreatic beta cells of these mice. Islets were isolated from these double transgenic mice and the fate of the cells once expressed insulin gene during the course of their differentiation was pursued by a confocal microscopy using EGFP expression as an indicator.
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