2000 Fiscal Year Final Research Report Summary
Construction of self microbe-protective transplantable epithelium by gene engineering
| Project/Area Number |
11650815
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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 |
生物・生体工学
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| Research Institution | Nagoya University |
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Principal Investigator |
SHINKAI Masashige Graduate School of Engineering, Nagoya University, Assistant Prof., 工学研究科, 助手 (70262889)
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| Co-Investigator(Kenkyū-buntansha) |
HATA Ken-ichiro Graduate School of Medicin, Nagoya University, Associate Prof., 医学研究科, 助教授 (80293710)
UEDA Minoru Graduate School of Medicin, Nagoya University, Prof., 医学研究科, 教授 (00151803)
KOBAYASHI Takeshi Graduate School of Engineering, Nagoya University, Prof., 工学研究科, 教授 (10043324)
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| Project Period (FY) |
1999 – 2000
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| Keywords | Gene therapy / transplantable epithelium sheet / Tissue engineering / Antimicrobial peptide activity / Sapecin / Epithelial cell / Cationic liposome / Mucosal cell |
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| Research Abstract |
In order to construct self microbe-protective transplantable epithelium, we investigated transfection method for epitherial cells or mucosal cells, plasmid construction of antimicrovial peptide gene, and its activity after transfection.
(1) Non-viral transfection method for oral mucosal cell was investigated. Six commercial transfection reagents were tested. The CellFECTIN performed best expression of transfected gene. Furthermore, the optimal condition of transfection was determined. When the mucosal cells were transfected at the optimal condition, the production of reporter protein increased up to ten times higher than other transfection reagents.
(2) A sapecin expression plasmid, pSecSape was constructed. Sapecin is one of antimicrobial peptide. The plasmid pSecSape was transfected in keratinocyte cells by our transfection method. Antimicrobial activity of supernatant of transfected keratinocyte cells was assessed by antimicrobial tests using E.coli, S.aureus, and P.aeruginosa. The supernatant strongly regressed these microbes.
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