Construction of self microbe-protective transplantable epithelium by gene engineering

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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.