| Project/Area Number |
14370544
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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 | 一般 |
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| Research Field |
Otorhinolaryngology
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| Research Institution | Yamaguchi University |
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Principal Investigator |
YAMASHITA Hiroshi Yamaguchi University, School of Medicine, Professor, 医学部, 教授 (00210419)
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| Co-Investigator(Kenkyū-buntansha) |
NAKAI Akira Yamaguchi University, Graduate School of Medicine, Professor, 大学院・医学研究科, 教授 (60252516)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥10,700,000 (Direct Cost: ¥10,700,000)
Fiscal Year 2004: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 2003: ¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2002: ¥5,700,000 (Direct Cost: ¥5,700,000)
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| Keywords | embryonic stem cell / organ culture / differentiation |
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| Research Abstract |
It was thought that hair cells of the inner ear of mammals do not regenerate after injury, although in birds and lower vertebrates hair cells regenerate spontaneously. Recently some studies about hair cell regeneration in mammals were reported using viruses, cell transplantation. We reported the co-culture of mouse cochlea with cells derived from embryoid bodies (EBs). EBs were obtained from mouse embryonic stem (ES) cells expressed green fluorescent protein (GFP). ES cells were differentiated to EBs in vitro using hanging drop technique. EBs were dissociated with trypsin-EDTA. Temporal bones were removed from 6d-old mice and picked up organ of cortis in PBS with 6 % glucose. They were incubated in DMEM F12 / 3g/l glucose/ N2supprement medium at 37℃ with 5 % CO_2 for 2 days. Then cells derived from EBs were added to cultured tissues and co-cultured for 3 days. For histological examination, cultured tissues were fixed with 4 % paraformaldehyde in PBS for 12 hours and stained sensory epithelium with phalloidin TexasRed. Then we observed cultured tissues under a fluorescent micro scope. Cultured organ of cortis for 7days were observed. They had same appearances compared with tissues observed immediately after removed so we thought that in this model organ cultures of corti were kept good condition for 7 days. The tissues co-cultured for 3 days were observed. Cells expressed GFP were attached to out side of sensory epithelium layer and no cells were identified in sensory epithelium layer.
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