Rivera-Tapia JA, Muñoz ZG, Román MC

Language: Spanish
References: 16
Page: 109-112
PDF size: 91.21 Kb.

ABSTRACT

Cell cultures can be contaminated with species belonging to the mycoplasma genus. Contaminating mycoplasmas may alter host cellular characteristics, for instance, alterations in growth characteristics, enzyme patterns, cell membrane composition and chromosomal abnormalities. These drawbacks emphasize the need to regularly screen cell cultures for mycoplasmal contamination. Objective: Mycoplasma detection in cell cultures came from several biomedical laboratories. Material and methods: Cells culture, were grown in absence of antibiotic and washed with PBS solution. Monolayer cells were first scraped off and suspended in 2 mL of SP-4 broth. Samples included in the microbiological culture were analyzed by PCR. Results: The microbiological study showed that 62.5% of the cultures was positive for the mycoplasma isolated and 37.5% negative. PCR technique determined the specific fragment (301 bp) for the mycoplasma genus. Conclusion: Because the mycoplasma frequently contaminate cell cultures, continuous detection is suggested, mainly in biomedical laboratories.

REFERENCES

1. Barile MF. Mycoplasma-tissue cell culture interactions. In: Tully JG, Whitcomb RF (ed). The mycoplasmas. New York: Academic Press, 1979; 425-474.

2. McGarrity GJ, Kotani H. Cell culture mycoplasmas. In: Razin S, Barile MF (ed). The mycoplasmas. New York: Academic Press, 1985; 353-390.

3. Stanbridge EJ. Mycoplasmas and cell cultures. Bacteriol Rev 1971; 35: 206-227.

4. Rivera-Tapia JA, Cedillo RL, Gil JC. Some biological features of Mollicutes. Rev Latinoam Microbiol 2002; 44: 53-57.

5. Del Giudice RA, Gardella RS, Hopps HE. Cultivation of formerly noncultivable strains of Mycoplasma hyorhinis. Curr Microbiol 1980; 4: 75-80.

6. Hopps HE, Meyer BC, Barile MF, Del Giudice RA. Problems concerning “non-cultivable” mycoplasma contaminants in tissue cultures. Ann NY Acad Sci 1973; 225: 265-276.

7. Gobel UB, Stanbridge E. Cloned mycoplasma ribosomal RNA genes for the detection of mycoplasma contamination of tissue cultures. Science 1984; 226: 1211-1213.

8. Johansson KE, Johansson I, Gobel UB. Evaluation of different hybridization in cell cultures. Mol Cell Probes 1990; 4: 33-42.

9. Razin S, Gross M, Wormser M, Pollack Y, Glaser G. Detection of mycoplasmas infecting cell cultures by DNA hybridization. In Vitro 1984; 20: 404-408.

10. Kuppeveld FJM, Johansson KE, Galama JMD, Kissing J, Bolske G, Van Der Logt JTM, Melchers WJG. Detection of mycoplasma contamination in cell cultures by a mycoplasma group-specific PCR. Appl Environ Microbiol 1994; 60: 149-152.

11. Spaepen M, Angulo AF, Marynen P, Cassiman JJ. Detection of bacterial and mycoplasma contamination in cell cultures by polymerase chain reaction. FEMS Microbiol Lett 1992; 99: 89-94.

12. Bolske G. Survey of mycoplasma infection in cell cultures and a comparison of detection methods. Zentralbl Bakteriol Mikrobiol 1988; Hyg A269: 331-340.

13. Shidu K, Rashidbaigi A, Testa D, Liao MJ. Competitor internal standards for quantitative detection of mycoplasma DNA. FEMS Microbiol Leet 1995; 128: 207-212.

14. Hay RJ. Operator induced contamination in cell culture systems. Dev Biol Stand 1991; 75: 193-204.

15. Lincoln CK, Gabridge MG. Cell culture contamination: Source, consequences, prevention and elimination. Methods Cells Biol 1998; 57: 49-65.

16. McGarrity GJ. Detection of mycoplasmas infection in cell cultures. Adv Cell Cult 1982; 2: 99-131.