2013 Fiscal Year Final Research Report

Development of a method to remove mycoplasmas from the contaminated cell cultures using antimicrobial photodynamic therapy

Research Project

Project/Area Number	23592692
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Multi-year Fund
Section	一般
Research Field	Morphological basic dentistry
Research Institution	Hokkaido University
Principal Investigator	HASEBE Akira 北海道大学, 歯学研究科(研究院), 助教 (90281815)
Co-Investigator(Kenkyū-buntansha)	柴田健一郎北海道大学, 大学院歯学研究科, 教授 (50145265)
Project Period (FY)	2011 – 2013
Keywords	光線力学療法 / マイコプラズマ汚染 / 培養細胞
Research Abstract	Mycoplasmas are one of the most frequent contaminants due to their characteristics in cell cultures. It is required to develop a method to remove mycoplasmas without using antibiotics, because a treatment with antibiotics may induce antibiotic-resistant bacteria. This study attempted to remove cell culture-contaminating Mycoplasma salivarium (Ms) by utilizing antimicrobial photodynamic therapy with a cordless red light emitting diode. Ms-infected human embryonic kidney 293 cells was irradiated with red light emitting diode in the presence of methylene blue as a photosensitizer, after decreasing the number of live Ms by washing with sterilized phosphate-buffered saline. No MS was detected in the contaminated cells by both mycoplasma detection kit and cultivation after antimicrobial photodynamic therapy, demonstrating that antimicrobial photodynamic therapy successfully remove Ms from its contaminated cells without giving damages to the cells.