The current industry standard for mycoplasma detection in biological products and cell cultures is specified by regulatory agencies (European Directorate for the Quality of Medicines, EDQM, Pharmakopoeia Europaea, PhEur; Center for Biologics Evaluation and Research, CBER, US Food and Drug Administration, FDA). The regulatory methods of choice include culturing methods on agar media directly, as well as inoculation of broth media with multiple subcultures on agar media. Cell culture-adapted mycoplasmas that are unable to grow on conventional media are detected by indicator cell procedures applying DNA-specific staining and fluorescence microscopy. Although the testing methods vary slightly between the different regulatory agencies, they are adequate for the sensitive detection of a broad spectrum of mycoplasma species known to contaminate cell cultures.