• Mysterious mycoplasma: a challenge for cell culture authentication.
  • Sajad Alimardan,1,* Ali Farhadi,2 Haniyeh Abuei,3 Hanie Arefzade,4 Sobhan Alimardan,5 Farshid Moosavi,6
    1. Department of Medical Biotechnology, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran
    2. Diagnostic Laboratory Sciences and Technology Research Center,School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran
    3. Department of Medical Biotechnology, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran
    4. Department of Medical Laboratory sciences, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran
    5. Department of Medical Laboratory sciences, School of Paramedical Sciences, Arak University of Medical Sciences, Arak, Iran
    6. Department of Medical Biotechnology, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran


  • Introduction: Cell cultures could be cross-contaminated with other cell lines or infected by microorganisms such as bacteria, viruses, and fungi. Mycoplasma infection is a major problem in cell culture. Mycoplasma can cause a wide range of inappropriate effects on cell cultures and increase concern about the application of cell cultures in cell therapy, biotechnological products and research. In this study, we provide applicable information about: 1. The biology, frequency and sources of mycoplasma species in cell cultures; 2. Consequences and effects of mycoplasma infection on the function of contaminated cell cultures; 3. Available techniques to identify mycoplasma contamination with focus on most authentic detection methods; 4. The best methods to inhibit mycoplasma contamination in cell cultures; 5. Recommended procedures and available methods for eradication of mycoplasmas from cell cultures.
  • Methods: In this review study, we included subject-related papers from international databases such as PubMed, Scopus and Google Scholar for articles published during 2002-2018. Out of 160 reviewed articles, 60 papers were ultimately selected. The keywords used were as follows: mycoplasma contamination, cell culture techniques, diagnosis, prevention and eradication of mycoplasma.
  • Results: According to these 60 selected papers, the prevalence of mycoplasma contamination varies from lab to lab, involving 10% to 85% of cell cultures. It is hard to eliminate mycoplasma infection in cell cultures, if not infeasible. Thus, the best methods for prevention and inhibition of infection transmission are: discarding the contaminated cultures, having a pre-set road map to examine cell cultures periodically by at least two trustworthy detection methods, such as DNA staining and PCR )the latter is based on the detection of 16S rRNA sequences of the most frequent contaminating species of mycoplasma(, and employing aseptic technique instructions for cell culture department personnel. However, antibiotics like Quinolones, BM-Cyclin, Plasmocin and Normocin may help to eradicate the contamination and purify valuable cells which are not possible to be replaced.
  • Conclusion: Due to the high rate of mycoplasma contamination in cell cultures, practical information on the sources, detection and elimination of mycoplasmas have been presented to decrease the incidence of contamination in order to get accurate results from researches and desirable product performance.
  • Keywords: Cell Culture Techniques, Mycoplasma contamination, Diagnosis, Prevention and Control.