| Project/Area Number |
16591764
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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 |
Ophthalmology
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| Research Institution | Dokkyo Medical University |
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Principal Investigator |
SENOO Tadashi Dokkyo University, School of Medicine, Ophthalmology, Professor, 医学部, 教授 (50206653)
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| Co-Investigator(Kenkyū-buntansha) |
CHIBA Keizo Dokkyo University, School of Medicine, Ophthalmology, Instructor, 医学部, 講師 (60146181)
TERADA Osamu Dokkyo University, School of Medicine, Ophthalmology, Assistant, 医学部, 助手 (50326881)
ISHIMARU Shinpei Dokkyo University, School of Medicine, Ophthalmology, Assistant, 医学部, 助手 (80406214)
OBARA Yoshitaka International University of Health and Welfare, School of Health Science, Professor, 保健学部, 教授 (70048354)
KIKUCHI Michiharu Dokkyo University, School of Medicine, Ophthalmology, Assistant, 医学部, 助手 (30382972)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2005: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2004: ¥800,000 (Direct Cost: ¥800,000)
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| Keywords | corneal endothelial cells / Cell Cycle-Associated Proteins / E2F family / 角膜内皮 / 培養増殖 / 細胞周期関連因子 / 細胞周期抑制因子 |
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| Research Abstract |
We studied that, in vivo these cells are arrested in G1-phase of cell cycle and are actively maintained in this non-replicative state.
Material and Methods : In this study, both human donor corneas and rabbit corneal endothelial cells were used. E2F family (E2F1, E2F2) were selected as transcription factor. Detection of cell cycle progression Cell cycle progression was detected by immunostaining for Ki67, a marker of actively cycling cell. 1) In this study, Condition of electroporator was as follows. Voltage : 25v, duration 20ms., Pulse interval : 100ms, repeat 8 times. 2)The transcription factor was overexpressed in corneal endothelial cells by transducting full length cDNA using electroporation.
Results and Consideration : It was examined that stimulate proliferation of corneal endothelial cells by overexpressing the transcription factor, E2F1 and E2F2. The structure and function of rabbit corneal endothelium could be restored after transiently inducing proliferation by overexpressing E2F1 and E2F2. Capability of rabbit corneal endothelial proliferation was increased 23% and 30% by overexpressing E2F1 and E2F2. In the human endothelial cells, Ki-67 positive cells were obtained. However, human corneal endothelial cells to reached M-phase were very low number. We suggested that these cells were arrested in either G2-phase, or induced apoptosis. We will determine whether decreased E2F expression and stimulated cell cycle progression stimulates apoptosis.
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