| Project/Area Number |
16591706
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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 |
Otorhinolaryngology
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
KITA Tomoko Kyoto University, Graduate School of Medicine, Researcher, 医学研究科, 客員研究員 (20362519)
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| Co-Investigator(Kenkyū-buntansha) |
NAKAGAWA Tkayuki Kyoto University, Graduate School of Medicine, Assistant Professor, 医学研究科, 助手 (50335270)
FUJINO Kiyohiro Kyoto University, Graduate School of Medicine, Assistant Professor, 医学研究科, 助手 (50359832)
HIRAUMI Harukazu Kyoto University, Graduate School of Medicine, Assistant Professor, 医学研究科, 助手 (10374167)
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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 |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2005: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2004: ¥3,100,000 (Direct Cost: ¥3,100,000)
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| Keywords | hair cell / embryonic stem cell / neurite interaction / neurotrophic factor / cell transplantation / gene transfer / 蝸牛有毛細胞 / 神経細胞 / 器官培養 / 神経ガイド因子 |
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| Research Abstract |
Present study advanced our initiative project of "Inner ear regeneration by the cell-transplantation" with a view to acquiring the regeneration of the auditory function. In the cell-transplantation studies, the effects of neurotrophic factors, especially of brain-derived neurotrophic factor (BDNF), on the interaction between hair cells and transplanted neural cells. Embryonic stem cell (ES cell)-derived neurons by SDIA treatment were used as transplants for cell therapy for the primary loss of spiral ganglion neurons.
In the in vitro study, ES cell-derived neurons were cocultured with auditory epithelium explants of P3 mice for a few days by the insert method. Vestibular sensory epithelia from neonatal mice were pretreated with papain. Embryonic stem cell-derived neurites were adjacent to or surrounding hair cells, and exhibited expression of synaptophysin. Examination under an electron microscope and whole-cell patch-clamp recordings revealed that the terminals of neurons formed synaps
… More
es, implying contact with auditory epithelium.
In the in vivo study, NIH3T3 cells were transfected with BDNF gene using lipofection and then transplanted into the mouse inner ear. Immunohistochemistry and western blotting demonstrated the survival of grafted cells in the cochlea for up to four weeks after transplantation. A BDNF-specific enzyme-linked immunosorbent assay revealed a significant increase in BDNF production in the inner ear following transplantation of engineered cells. These findings indicate that cell-gene therapy with non-viral vectors may be applicable for the local, sustained delivery of therapeutic molecules into the cochlea. Mouse ES cell-derived neurons were implanted into the base of the cochlear modiolus of normal or deafened guinea pigs, which contains spiral ganglion neurons and cochlear nerve fibers. Both histological and functional assessments indicated the potential of transplanted cells to elicit the functional recovery of damaged cochleae.
These findings suggest the potential of the cell-gene therapy for the inner ear regeneration. Less
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