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Prolonged hyperthermia eliminates mycoplasma from cultured human and rat brain tumor cell lines
1Department of Neurological Surgery, School of Medicine, University of California, San Francisco 94143-0520.
Abstract:
Mycoplasma infection of mammalian cells in culture is a common occurrence that can affect the results of experimental protocols. Current methods of eliminating mycoplasma from cell cultures are usually tedious, time-consuming, and sometimes unsuccessful. In the present study, four cultured brain tumor cell lines (human U-251 MG, U-87 MG, SF-126, and rat 9L) were heavily contaminated with Mycoplasma orale. Heating the cultures to 41 degrees C for at least 96 h eliminated the contamination for up to 7 months, the maximum period of observation. The time chosen to assay for the presence of mycoplasma in cultures was critical: in some cultures heated for less than 96 h that initially appeared to be free of contamination, mycoplasma began to appear after 2 weeks. Heat-treated cells grew at the same rate as unheated control cells. Infected cells were more sensitive to X rays than uncontaminated cells, but the sensitivity reverted to normal after mycoplasma was eliminated by hyperthermia. The heating method does not require a cell cloning procedure or the use of exogenous materials. Treated cell cultures exhibit normal growth and radiation sensitivity, and the technique seems to be reliable and efficient.
Insights
Hyperthermia treatment effectively eliminates Mycoplasma contamination in cell cultures. This 41°C heat treatment for 96 hours offers a reliable method for mycoplasma removal, restoring normal cell growth and radiation sensitivity.
Area of Science:
- Cell Biology
- Microbiology
- Biotechnology
Background:
- Mycoplasma contamination is a frequent issue in mammalian cell cultures, potentially compromising experimental outcomes.
- Existing mycoplasma elimination methods are often inefficient, laborious, and time-consuming.
Purpose of the Study:
- To evaluate the efficacy of hyperthermia as a method for eradicating Mycoplasma orale contamination in cultured brain tumor cell lines.
- To assess the impact of hyperthermia on cell growth rate and radiation sensitivity.
Main Methods:
- Four brain tumor cell lines (human U-251 MG, U-87 MG, SF-126, and rat 9L) were intentionally infected with Mycoplasma orale.
- Cultures were subjected to heat treatment at 41°C for a minimum of 96 hours.
- Cell growth rates and radiation sensitivity were monitored post-treatment.
Main Results:
- Heating cultures at 41°C for at least 96 hours successfully eliminated Mycoplasma contamination for up to 7 months.
- A critical observation period of 96 hours was established, as shorter durations led to re-contamination.
- Heat-treated cells demonstrated normal growth rates and radiation sensitivity comparable to uninfected controls.
Conclusions:
- Hyperthermia at 41°C for 96 hours is an effective, efficient, and reliable method for eliminating Mycoplasma contamination in brain tumor cell lines.
- This technique does not require cell cloning or exogenous agents and preserves normal cell characteristics.
- The study presents a practical solution for researchers dealing with mycoplasma-infected cell cultures.