| Project/Area Number |
05671436
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Otorhinolaryngology
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| Research Institution | Kagoshima University |
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Principal Investigator |
HANAMURE Yutaka Kagoshima University, University Hospital, Assistant Professor, 医学部附属病院, 講師 (40145496)
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| Co-Investigator(Kenkyū-buntansha) |
FUKUDA Katsunori Kagoshima University, University Hospital, Assistant Professor, 医学部附属病院, 講師 (90156779)
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| Project Period (FY) |
1993 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1995: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1994: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1993: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Keywords | Respiratory epithelial cell / Differentiation / Ciliated cell / Secretory cell / Monoclonal antibody / Sialic acid / Interleukin-1beta / Ciliogenesis / 繊毛新生 / 繊毛運動 / 単クローン抗体 / 培養 / シアル酸 |
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| Research Abstract |
(1) We demonstrated a floating culture model which allowed effective induction of cell differentiation of human respiratory epithelial cells in vitro. Human nasal epithelial cells plated on type I collagen gel rapidly proliferated to reach confluence and showed flat squamous-like appearance, and after detached and floated gradually differentiate to secretory cells and ciliated cells and then displayd pseudostratified mucociliary epithelium after 14 days in floating culture. Ultrastructural and morphometric studies demonstrated fibrous granules, deuterosomes, and centrioles, the intracellular characteristic of ciliogenesis during floating and final high density of ciliated cells.
(2) Surface of matured ciliated cells are covered with glycocaryx that contains sialic acid as terminal carbohydrate of glycoconjugates. We used lectins, Macckia amurensis agglutinin and Sambucus nigra agglutinin which recognize sialic acid residues to observe expression of sialic acid residue during respiratory cell differentiation. Sialic acid was not present before start of ciliogenesis, gradually appeared during ciliogenesis and finally covered the full surface of matured ciliated cells. This result indicates that sialic acid is one of markers of respiratory cell differentiation.
(3) A novel monoclonal antibody, MAb-RPCS was obtained after immunization of mice with whole cells of 3 days floating eulture. MAb-RPCS recognized the secretory cells appeared at preciliated state. Western blotting with periodate oxidation revealed that MAb-RPCS recognized carbohydrate moieties.
(4) Interleukin-1beta (IL-1beta) is one of inflammatory cytokines that modulates inflammatory process. We observed the effects of IL-1beta during the differentiation of respiratory epithelial cells. The results suggest that IL-1beta accelerates the appearance of secretory cells and suppresses the onset of ciliogenesis and the appearance of matured ciliated cells.
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