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

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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.