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Treatment of mycoplasma-contaminated continuous cell lines with mycoplasma removal agent (MRA)
Abstract:
Thirty-nine continuous adherent or suspension cell lines were treated with a quinolone antibiotic, Mycoplasma Removal Agent (MRA), for the elimination of chronic mycoplasma contamination. In preliminary experiments MRA did not show any cytostatic or cytotoxic effects on mycoplasma-free cell cultures in concentrations up to ten-fold the concentration used for mycoplasma eradication. Twenty-eight cell lines (72%) were effectively cleansed of the mycoplasma contaminants by MRA treatment. The persistent removal of the mycoplasma infection was monitored by three mycoplasma detection assays. In seven cell lines (18%) the mycoplasmas were resistant to treatment with MRA. The resistant species was mainly M. arginini followed by M. orale and A. laidlawii; however, other cell lines harboring these species were cured. Four cell lines (10%) which prior to treatment presented with decreased viability and poor or no cell growth were lost during or shortly after the exposure to the antibiotic. If an antibiotic elimination is attempted it is imperative to closely examine the effectiveness of treatment and possible eukaryotic cytotoxicity. The treated mycoplasma-free cells may also no longer express the original features as a result of treatment or the absence of mycoplasma.
Insights
Mycoplasma Removal Agent (MRA) effectively eradicated mycoplasma contamination in 72% of cell lines. However, some cell lines showed resistance or loss of viability, necessitating careful monitoring during antibiotic treatment for mycoplasma removal.
Area of Science:
- Cell biology
- Microbiology
- Pharmacology
Background:
- Chronic mycoplasma contamination is a common issue in cell cultures.
- Mycoplasma contamination can significantly impact cell behavior and experimental results.
- Effective elimination of mycoplasma is crucial for reliable cell-based research.
Purpose of the Study:
- To evaluate the efficacy of Mycoplasma Removal Agent (MRA) in eliminating chronic mycoplasma contamination from cell lines.
- To assess the safety and potential side effects of MRA treatment on eukaryotic cell cultures.
- To identify mycoplasma species resistant to MRA treatment.
Main Methods:
- Treatment of 39 adherent or suspension cell lines with MRA.
- Preliminary assessment of MRA's cytostatic and cytotoxic effects on mycoplasma-free cells.
- Monitoring mycoplasma eradication using three distinct detection assays.
- Evaluation of cell viability and growth post-treatment.
Main Results:
- MRA successfully eradicated mycoplasma in 28 out of 39 (72%) cell lines.
- Mycoplasma resistance to MRA was observed in 7% (18%) of cell lines, primarily M. arginini, M. orale, and A. laidlawii.
- Four cell lines (10%) were lost due to decreased viability or growth inhibition during or after MRA treatment.
- MRA showed no significant cytostatic or cytotoxic effects on mycoplasma-free cells at effective eradication concentrations.
Conclusions:
- MRA is an effective agent for eliminating mycoplasma contamination in a majority of cell lines.
- Careful monitoring for treatment effectiveness and potential eukaryotic cytotoxicity is essential when using MRA.
- Cell line characteristics may be altered post-treatment or due to mycoplasma absence.