Accumulating data implicate mycoplasma contamination as the single biggest problem in the culture of continuous cell lines. Mycoplasma infection can affect virtually every parameter and functional activity of the eukaryotic cells. A successfull alternative to discarding infected cultures is to attempt to eliminate the contaminants by treatment with specific and efficient antimycoplasma antibiotics. The addition of antibiotics to the culture medium during a limited period of time (1–3 wk) is a simple, inexpensive, and very practical approch for decontaminating continuous cell lines. Here, we examined the effectiveness of several antibiotic treatment protocols that we have employed routinely in our cell lines bank. On an aggregate, 673 cultures from 236 chronically mycoplasma-positive cell lines were exposed to one of the following five antibiotic regimens: mycoplasma removal agent (quinolone; a 1-wk treatment), enrofloxacin (quinolone; 1 wk), sparfloxacin (quinolone; 1 wk), ciprofloxacin (quinolone; 2 wk), and BM-Cyclin (alternating tiamulin and minocycline; 3 wk). The mycoplasma infection was permanently (as determined by three solid mycoplasma detection assays) eliminated by the various antibiotics in 66–85% of the cultures treated. Mycoplasma resistance was seen in 7–21%, and loss of the culture as a result of cytotoxically caused cell death occurred in 3–11% of the cultures treated. Overall, 223 of the 236 mycoplasma-positive cell lines could be cured in a first round of antibiotic treatment with at least one regimen. Taken together, 95% of the mycoplasma-infected cell lines were permanently cleased of the contaminants by antibiotic treatment, which validates this approach as an efficient and technically simple mycoplasma eradication method.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Barile, M. F. Mycoplasma-tissue cell interactions. In: Tully, J. G.; Whitcomb, R. F., ed. The mycoplasmas. Vol. 2. New York: Academic Press; 1979;425–474.
Drexler, H. G., ed. The leukemia-lymphoma cell line factsbook. San Diego, CA: Academic Press; 2000.
Drexler, H. G.; Dirks, W. G.; MacLeod, R. A. F. False human hematopoietic cell lines: cross-contaminations and misinterpretations. Leukemia 13:1601–1607; 1999.
Drexler, H. G.; Dirks, W. G.; MacLeod, R. A. F.; Quentmeier, H.; Steube, K. G.; Uphoff, C. C., ed. DSMZ catalogue of human and animal cell lines. 8th ed. Braunschweig, Germany: DSMZ; 2001.
Drexler, H. G.; Gignac, S. M.; Hu, Z. B.; Hopert, A.; Fleckenstein, E.; Voges, M.; Uphoff, C. C. Treatment of mycoplasma contamination in a large panel of cell cultures. In Vitro Cell. Dev. Biol. 30A:344–347; 1994.
Drexler, H. G.; Uphoff, C. C. Contamination of cell culture, mycoplasma. In: Spier, E., et al., ed. The encyclopedia of cell technology. New York: J. Wiley; 2000:609–627.
Fleckenstein, E.; Uphoff, C. C.; Drexler, H. G. Effective treatment of mycoplasma contamination in cell lines with enrofloxazin (Baytril). Leukemia 8:1424–1434; 1994.
Gignac, S. M.; Brauer, S.; Häne, B.; Quentmeier, H.; Drexler, H. G. Elimination of mycoplasma from infected leukemia cell lines. Leukemia 5:162–165; 1991.
Gignac, S. M.; Uphoff, C. C.; MacLeod, R. A. F.; Steube, K.; Voges, M.; Drexler, H. G. Treatment of mycoplasma-contaminated continuous cell lines with mycoplasma removal agent (MRA). Leukemia Res. 16:815–822; 1992.
Hay, R. J.; Macy, M. L.; Chen, T. R. Mycoplasma infection of cultured cells. Nature 339:487–488; 1989.
MacLeod, R. A. F.; Dirks, W. G.; Kaufmann, M.; Matsuo, Y.; Milch, H.; Drexler, H. G. Widespread intraspecies cross-contamination of human tumor cell lines arising at source. Int. J. Cancer 83:555–563: 1999.
Uphoff, C. C.; Brauer, S.; Grunicke, D., et al. Sensitivity and specificity of five different mycoplasma detection assays. Leukemia 6:335–341; 1992a
Uphoff, C. C.; Drexler, H. G. Comparative PCR analysis for detection of mycoplasma infections in continuous cell lines. In Vitro Cell. Dev. Biol. 38:79–85, 2002.
Uphoff, C. C.; Gignac, S. M., Drexler, H. G. Mycoplasma contamination in human leukemia cell lines. I. Comparison of various detection methods. J. Immunol. Methods 149:43–53; 1992b.
Uphoff, C. C.; Gignac, S. M.; Drexler, H. G. Mycoplasma contamination in human leukemia cell lines. II. Elimination with various antibiotics. J. Immunol. Methods 149:55–62, 1992c.
Vollenbroich, D.; Pauli, G.; Özel, M.; Vater, J. Antimycoplasma properties and application in cell culture of surfactin, a lipopeptide antibiotic from Bacillus subtilis. Appl. Environ. Microbiol. 63:44–49; 1997.
About this article
Cite this article
Uphoff, C.C., Drexler, H.G. Comparative antibiotic eradication of mycoplasma infections from continuous cell lines. In Vitro Cell.Dev.Biol.-Animal 38, 86–89 (2002). https://doi.org/10.1290/1071-2690(2002)038<0086:CAEOMI>2.0.CO;2
- cell culture