Mycoplasma contamination in human leukemia cell lines. II: Elimination with various antibiotics

19 suspension cell lines were treated with antibiotics for elimination of chronic contamination with mycoplasma. We compared the efficiency, cytotoxicity and cross-resistance of the commercially available antibiotics MRA (Mycoplasma Removal Agent, a quinolone derivative and DNA gyrase inhibitor), Ciprobay (ciprofloxacin, also a quinolone derivative and DNA gyrase inhibitor), and BM-cyclin (a combination of tiamulin, a pleuromutilin derivative, and minocycline, a tetracycline derivative, both inhibitors of protein synthesis on ribosomes). Contaminants were eliminated in all 19 cell lines by BM-Cyclin. Only 74% of the cell lines were cleared of contamination by both MRA and Ciprobay. Successful treatment was monitored by three mycoplasma detection assays. Cross-resistance was noted between MRA and Ciprobay in four of the five cell lines not cleared by either reagent. This resistance could, however, be overcome by consecutive exposure to BM-cyclin. Employed at the recommended concentrations, the antibiotics did not cause marked cytotoxicity, but the growth of the cells was affected to various degrees by some antibiotics. The elimination of mycoplasma from chronically contaminated cell lines is an effective alternative to other treatment protocols, but is cost-intensive and time-consuming; lasting damaging effects of the treatments on the eukaryotic cells cannot be excluded. Long-term post-treatment monitoring is mandatory, since contaminants may only be suppressed and then recur.