Elimination of mycoplasmas from cell cultures by a novel soft agar technique

Abstract

Mycoplasmal infection of cell cultures remains a significant threat to diagnostic and research procedures. In certain defined situations, curing of mycoplasmal infected cultures is a reasonable exercise. Four methods of curing were compared: treatment with BM-cycline, 5 bromouracil, use of specific antisera and treatment of infected cells suspended in soft agar with antibiotics. Antisera treatments were of low efficiency of curing: 50%. None of nine infected cell lines treated with 5-bromouracil were consistently cured of mycoplasmas. The use of BM-cycline was effective for some, but not all lines and required long periods of treatment, 12-21 days. 35 naturally or deliberately infected cultures were treated in soft agar a total of 119 times. This procedure which consisted of suspending infected cultures in soft agar containing appropriate antibiotics resulted in successful mycoplasmal elimination 118/119 times. This soft agar technique took 1-3 days. In separate studies, it was shown that certain Mycoplasma fermentans strains were resisted to this and other curing methods. This may be due to their intracellular location. Such strains may be more amenable to antibiotics that penetrate mammalian cells. It is concluded that the soft agar technique is a rapid, efficient and reliable method to eliminate cell culture mycoplasmas.

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

“…A number of decontamination protocols have been employed in recent years with different efficiencies (Lincoln and Lundin 1990;Kotani et al 1991;Fleckenstein and Drexler 1996). However, the most promising technique, which is relatively simple and inexpensive, appears to be antibiotic treatment (Fleckenstein and Drexler 1996).…”

Effect of Antibiotics against Mycoplasma sp. on Human Embryonic Stem Cells Undifferentiated Status, Pluripotency, Cell Viability and Growth

“…As a consequence, mycoplasmal infection of cell cultures could often persist for long periods of time without being noticed and with no apparent cell damage [9]. However, contaminating mycoplasmas affect virtually every parameter within the cell culture system.…”