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Eliminating mycoplasmas from contaminated cell cultures
Israel Journal of Medical Sciences
|July 1, 1981
Summary
A new method selectively kills mycoplasma contaminants in cell cultures by incorporating 5-bromouracil into their DNA, making it susceptible to light-induced breaks. This technique effectively eliminates mycoplasma contamination from mammalian cell lines.
Area of Science:
- Cell Biology
- Microbiology
- Molecular Biology
Background:
- Mycoplasmas are common and problematic contaminants in cell cultures.
- Existing methods for mycoplasma elimination are often ineffective or lack simplicity and speed.
Purpose of the Study:
- To develop a simple, rapid, and effective method for selectively eliminating mycoplasma contamination from mammalian cell cultures.
Main Methods:
- Exploited differences in nucleic acid metabolism between mycoplasmas and host cells.
- Utilized selective incorporation of 5-bromouracil (5-BrUra) into mycoplasma DNA.
- Employed photosensitization with 33258-Hoechst fluorochrome and visible light to induce DNA breaks.
Main Results:
- Demonstrated selective killing of Mycoplasma hyorhinis and Acholeplasma laidlawii in Chinese hamster cells.
- Successfully obtained mycoplasma-free cell clones.
- Showed efficacy in curing BHK-21 and RAG cells from unknown mycoplasma strains.
Conclusions:
- The developed method offers a generally applicable approach for eliminating mycoplasma contamination from various mammalian cell cultures.
- The technique leverages the unique A+T rich DNA content of mycoplasmas for targeted eradication.