Selective killing of mycoplasmas from contaminated mammalian cells in cell cultures

Advances in PCR-based detection of mycoplasmas contaminating cell cultures.

“…It is often better to destroy the infected culture by autoclaving and replacing it with a mollicute-free stock culture. However, when the infected cell culture is irreplaceable, the following procedures can be used to eliminate mollicutes: (7'1~ treatment with anti-mollicute antisera, passage through mice, exposure to murine macrophages, treatment of cells with 5-bromouracil or merocyanine 540 and Hoechst 33258 followed by exposure to light, (68,69) prolonged heat treatment, and selective killing with antibiotics [tylosin, spectinomycin, tiamulin, minocyclin, gentamycin, neomycin, kanamycin, novobiocin, thialine, quinolones, MRA (ICN), BM-Cyclin (Boerhinger Mannhiem), etc. ], or mycoplasmacidal drugs.…”

Contamination of Tissue Cultures by Mycoplasmas

“…Eliminating mycoplasmas by passage of a cell culture through nude mice [109] has been successful for some, but not all, mycoplasmas. An efficient procedure for eliminating mycoplasmas is based on the selective incorporation of 5-bromouracil (5-BrUra) into mycoplasmas, and the induction of breaks by light in the 5-BrUra-containing DNA [110]. The unusually high content of A+T makes the mycoplasma DNA an excellent candidate for the induction of breakage by the combined action of 5-BrUra, 33258-Hoechst and visible light [110].…”

“…An efficient procedure for eliminating mycoplasmas is based on the selective incorporation of 5-bromouracil (5-BrUra) into mycoplasmas, and the induction of breaks by light in the 5-BrUra-containing DNA [110]. The unusually high content of A+T makes the mycoplasma DNA an excellent candidate for the induction of breakage by the combined action of 5-BrUra, 33258-Hoechst and visible light [110]. Some of the elimination procedures may apply to some, but not all, mycoplasma species; some of them are laborious and/or time consuming.…”

Some properties and applications of monoclonal antibodies

“…Another problem is contamination with Mvcoplasma, which causes cultures to die out gradually. It appears to be controllable by injecting infected cells into mice as for the production of ascites (Hudson & Hay, 1980) and recovering the apparently Mvcoplasma-free cells when the ascites is drained (P. A. W. Edwards, unpublished work; see also the methods of Marcus et al, 1980 andSchimmelpfeng et al, 1980). Apparent instability of hybrids, i.e.…”