Current Issue

Volume 21, Number 2, Jul-Sep (Summer) 2019 , Serial Number: 82 Pages: 143-149

Effectiveness of Plasmocure™ in Elimination of Mycoplasma Species from Contaminated Cell Cultures: A Comparative Study versus Other Antibiotics


Vahid Molla Kazemiha, M.Sc, 1, Shahram Azari, Ph.D, 1, Mahdi Habibi-Anbouhi, Ph.D, 1, Amir Amanzadeh, Ph.D, 1, Shahin Bonakdar, Ph.D., 1, Mohammad Ali Shokrgozar, Ph.D., 1, *, Reza Mahdian, M.D., Ph.D., 2, *,
National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran
Department of Molecular Medicine, Pasteur Institute of Iran, Tehran, Iran
*Corresponding Addresses: P.O.Box: 1316943551 National Cell Bank of Iran Pasteur Institute of Iran Tehran Iran P.O.Box: 1316943551 Department of Molecular Medicine Pasteur Institute of Iran Tehran Iran Emails:mashokrgozar@pasteur.ac.ir,dr.reza.mahdian@gmail.com

Abstract

Objective

Mycoplasmas spp. is among major contaminants of eukaryotic cell cultures. They cause a wide range of problems associated with cell culture in biology research centers or biotechnological companies. Mycoplasmas are also resistant to several antibiotics. Plasmocin™ has been used to treat cell lines but Plasmocin™-resistant strains have been reported. InvivoGen has developed a new anti-Mycoplasma agent called Plasmocure™ in order to eliminate resistant Mycoplasma contamination. The aim of this study was the selection of the best antibiotics for treatment of mycoplasma in cell cultures.

Materials and Methods

In this experimental study, a total of 100 different mammalian cell lines contaminated with different Mycoplasma species were evaluated by microbiological culture (as the gold standard method), indirect DNA fluorochrome staining, enzymatic (MycoAlert™), and universal or species-specific polymerase chain reaction (PCR) detection methods. In this study, animal and human cell lines available in National Cell Bank of Iran, were treated with Plasmocure™. The treatment efficacy and cytotoxicity of Plasmocure™ were compared with those of commonly used antibiotics such as BM- cyclin, Plasmocin™, MycoRAZOR™, sparfloxacin and enrofloxacin.

Results

Plasmocure™ is comprised of two antibiotics that act through various mechanisms of action than those in Plasmocin™. Two-week treatment with Plasmocure™ was enough to completely eliminate Mycoplasma spp. A moderate toxicity was observed during Mycoplasma treatment with plasmocure™; But, after elimination of Mycoplasma, cells were fully recovered. Mycoplasma infections were eliminated by Plasmocure™, BM-cyclin, Plasmocin™, MycoRAZOR™, sparfloxacin and enrofloxacin. However, the outcome of the treatment process (i.e. the frequency of complete cure, regrowth or cell death) varied among different antibiotics.

Conclusion

The highest number of cured cell lines was achieved by using Plasmocure™ which also had the lowest regrowth rate after a period of four months. As a conclusion; Plasmocure™ might be considered an effective antibiotic to treat Mycoplasma infections in mammalian cell cultures especially for precious or vulnerable cells.