Skip to main content

Advertisement

Log in

PCR-based detection and eradication of mycoplasmal infections from various mammalian cell lines: a local experience

  • Original Research
  • Published:
Cytotechnology Aims and scope Submit manuscript

Abstract

A total of 200 cell lines including different human, monkey, mice, hamster and rat cell types were examined for mycoplasma infection status. PCR assay using generic-specific universal primers showed that 40 (20%) of the cell lines are contaminated with mycoplasma. Employment of species-specific primers within these infected cell lines revealed infection with M. hyorhinis (42.5%), M. fermentas (37.5%), M. arginini (37.5%), M. orale (12.5%) and A. laidlawii (7.5%). A number of the cultures were coinfected with 2 or 3 different species. Contaminated samples were treated with BM-Cyclin, Ciprofloxacin and mycoplasma removal agent (MRA). Mycoplasma eradication was subsequently checked by PCR following 2 weeks continuous culture of treated cells in antibiotic free culture medium. Mycoplasmal infections were eradicated in 100, 70 and 42% of infected cell lines when the samples were treated with BM-Cyclin, MRA and Ciprofloxacin, respectively. However, 12% (BM-Cyclin), 62.5% (MRA) and 82.5% (Ciprofloxacin) of mycoplasma regrowth was observed 4 months after the treatment. Notably, the risk of spontaneous culture death was 17.5, 12.5 and 0% for BM-Cyclin, MRA and Ciprofloxacin, respectively.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1

Similar content being viewed by others

References

  • Barile MF, Rottem S (1993) Mycoplasma in cell culture. In: Kahane Ι, Adoni A (eds) Rapid diagnosis of mycoplasmas. Plenum Press, New York, pp 155–193

    Google Scholar 

  • Blanchard A, Montagnier L (1994) AIDS-associated mycoplasma. Annu Rev Microbiol 48:687–712

    Article  CAS  Google Scholar 

  • Boolske G (1988) Survey of mycoplasma in cell cultures and a comparison of detection methods. Zentralbl Bakteriol Mikrobiol Hyg 269:331–340

    Google Scholar 

  • Drexler HG, Uphoff CC (2002) Mycoplasma contamination of cell cultures: incidence, sources, effects, detection, elimination, prevention. Cytotechnology 39:75–90

    Article  CAS  Google Scholar 

  • Drexler HG, Dirks W, Macleod RAF et al (2001) DSMZ catalogue of human and animal cell lines, 8th edn. DSMZ, Braunschweig

    Google Scholar 

  • Drexler HG, Uphoff CC, Dirks WG et al (2002) Mix-ups and mycoplasma: the enemies within. Leuk Res 26:329–333

    Article  Google Scholar 

  • Fleckenstein E, Drexler HG (1996) Elimination of mycoplasma contamination in cell cultures. Biochemica 1:48–51

    Google Scholar 

  • Fleckenstein E, Uphoff CC, Drexler HG (1994) Effective treatment of mycoplasma contamination in cell lines with enrofloxacin (Baytril). Leukemia 8:1424–1434

    CAS  Google Scholar 

  • Gardella RS, Del Guidice RA (1984) Antibiotic sensitivities and elimination of mycoplasmas from infected cell cultures. Isr J Med Sci 20:931–934

    CAS  Google Scholar 

  • Gurbay A, Gontheir B, Barret L et al (2007) Cytotoxic effect of ciprofloxacin in primary culture of rat astrocytes and protection by vitamin E. Toxicology 229:54–61

    Article  Google Scholar 

  • Hopert A, Uphoff CC, Wirth M et al (1993) Specificity and sensitivity of polymerase chain reaction (PCR) in comparison with other methods for the detection of mycoplasma contamination in cell lines. J Immunol Meth 164:91–100

    Article  CAS  Google Scholar 

  • Jung H, Wang SY, Ding WH et al (2003) Detection and treatment of mycoplasma contamination in cultured cells. Chang Gung Med J 26:250–258

    Google Scholar 

  • Kong F, James G, Gordon S et al (2001) Species-specific PCR for identification of common contaminant Mollicutes in cell culture. Appl Environ Microbiol 67:3195–3200

    Article  CAS  Google Scholar 

  • Kotani H, Butler G, Heggan D et al (1991) Elimination of mycoplasma from cell cultures by a novel soft agar technique. In Vitro Cell Dev Biol 27A:509–513

    Article  CAS  Google Scholar 

  • Lincoln CK, Lundin DJ (1990) Mycoplasma detection and control. US FCC Newsl 20:1–3

    Google Scholar 

  • Mattsson JG, Johansson KE (1993) Oligonucleotide probes complementary to 16S rRNA for rapid detection of mycoplasma contamination in cell cultures. FEMS Microbiol Lett 107:139–144

    Article  CAS  Google Scholar 

  • Razin S, Yogev D, Naot Y (1998) Molecular biology and pathogenicity of mycoplasma. Microbiol Mol Biol Rev 62:1094–1156

    CAS  Google Scholar 

  • Robinson LB, Wichelhausen RH (1956) Contamination of human cell cultures by pleuropneumonia like organisms. Science 124:1147–1148

    Article  CAS  Google Scholar 

  • Rottem S, Barile MF (1993) Beware of mycoplasma. Trends Biotechnol 11:143–151

    Article  CAS  Google Scholar 

  • Rottem S, Naot Y (1998) Subversion and exploitation of host cells by mycoplasmas. Trends Microbiol 6:436–440

    Article  CAS  Google Scholar 

  • Somasundaram C, Matzku S, Nickas W (1992) Use of ciprofloxacin and BM-Cyclin in mycoplasma decontamination. In Vitro Cell Dev Biol 28A:708–710

    Article  CAS  Google Scholar 

  • Tang J, Hu M, Lee S et al (2000) A polymerase chain reaction based method for detecting Mycoplasma/Acholeplasma contaminants in cell culture. J Microbial Methods 39:121–126

    Article  CAS  Google Scholar 

  • Timenetsky J, Santos L, Buzinhani M et al (2006) Detection of multiple mycoplasma infection in cell cultures by PCR. Braz J Med Biol Res 39:907–914

    Article  CAS  Google Scholar 

  • Uphoff CC, Drexler HG (2002) Comparative PCR analysis for detection of mycoplasma infections in continuous cell lines. In Vitro Cell Dev Bio Anim 38:79–85

    Article  CAS  Google Scholar 

  • Uphoff CC, Gignac SM, Drexler HG (1992) Mycoplasma contamination in human leukemia cell lines. П. Elimination with various antibiotics. J Immunol Methods 149:55–62

    CAS  Google Scholar 

  • Uphoff CC, Meyer C, Drexler HG (2002) Elimination of mycoplasma from leukemia-lymphoma cell lines using antibiotics. Leukemia 16:284–288

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Fazel Shokri.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Molla Kazemiha, V., Shokrgozar, M.A., Arabestani, M.R. et al. PCR-based detection and eradication of mycoplasmal infections from various mammalian cell lines: a local experience. Cytotechnology 61, 117–124 (2009). https://doi.org/10.1007/s10616-010-9252-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10616-010-9252-6

Keywords

Navigation