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Treatment of mycoplasma contamination in a large panel of cell cultures
H G Drexler1, S M Gignac, Z B Hu
1DSM-German Collection of Microorganisms and Cell Cultures, Department of Human and Animal Cell Cultures, Braunschweig, Germany.
Abstract:
Mycoplasmal contamination remains a significant impediment to the culture of eukaryotic cells. For certain cultures, attempts to eliminate the infection are feasible alternatives to the normally recommended disposal of the contaminated culture. Here, three antibiotic regimens for mycoplasmal decontamination were compared in a large panel of naturally infected cultures: a 1-wk treatment with the fluoroquinolone mycoplasma removal agent (MRA), a 2-wk treatment with the fluoroquinolone ciprofloxacin, and three rounds of a sequential 1-wk treatment with BM-Cyclin containing tiamulin and minocyclin. These antibiotic treatments had a high efficiency of permanent cure: MRA 69%, ciprofloxacin 75%, BM-Cyclin 87%. Resistance to mycoplasma eradication was observed in some cell cultures: BM-Cyclin 0%, MRA 20%, ciprofloxacin 20%. Nearly all resistant contaminants that could be identified belonged to the species Mycoplasma arginini and M. orale. Detrimental effects of the antibiotics were seen in the form of culture death caused by cytotoxicity (in 5 to 13% of the cultures). Alterations of the cellular phenotypic features or selective clonal outgrowth might represent further untoward side effects of exposure to these antibiotics. Overall, antibiotic decontamination of mycoplasmas is an efficient, inexpensive, reliable, and simple method: 150/200 (75%) chronically and heavily contaminated cultures were cured and 50/200 (25%) cultures could not be cleansed and were either lost or remained infected. It is concluded that eukaryotic cell cultures containing mycoplasmas are amenable to antibiotic treatment and that a cure rate of three-quarters is a reasonable expectation.
Insights
Antibiotic treatments effectively decontaminate mycoplasma-infected eukaryotic cell cultures, with BM-Cyclin showing the highest cure rate (87%). While some resistance occurred, this method offers a reliable alternative to discarding cultures.
Area of Science:
- Cell Biology
- Microbiology
- Pharmacology
Background:
- Mycoplasmal contamination is a major challenge in eukaryotic cell culture.
- Disposal of contaminated cultures is standard, but decontamination is sometimes feasible.
- Effective antibiotic regimens are needed to combat mycoplasma infections.
Purpose of the Study:
- To compare the efficacy of three antibiotic regimens for mycoplasma decontamination in cell cultures.
- To assess cure rates, resistance, and potential adverse effects of different treatments.
- To determine the overall viability of antibiotic decontamination as a strategy.
Main Methods:
- Three antibiotic treatments were tested: mycoplasma removal agent (MRA, 1 week), ciprofloxacin (2 weeks), and BM-Cyclin (3 rounds of 1 week).
- Treatments were applied to naturally infected cell cultures.
- Cure rates, resistance patterns (identifying species like Mycoplasma arginini and M. orale), and cytotoxic effects were evaluated.
Main Results:
- BM-Cyclin achieved the highest cure rate (87%), followed by ciprofloxacin (75%) and MRA (69%).
- Resistance to eradication was observed, particularly with MRA and ciprofloxacin (20% each), while BM-Cyclin showed no resistance.
- Cytotoxicity led to culture death in 5-13% of cases, with potential for phenotypic alterations.
Conclusions:
- Antibiotic decontamination is an efficient, cost-effective, and reliable method for treating mycoplasma-contaminated eukaryotic cell cultures.
- A significant proportion (75%) of contaminated cultures can be successfully cured.
- Eukaryotic cell cultures are amenable to antibiotic treatment, with a three-quarters cure rate being a reasonable expectation.