Detection of mycoplasma contamination through modulation (stimulation or inhibition) of thymidine incorporation by unstimulated mouse spleen cells.
In the present report we describe a rapid and sensitive assay for mycoplasma detection in cell cultures. The assay is based on the ability of contaminated culture supernatants to modulate [3H]TdR incorporation by unstimulated mouse splenocytes. Several mycoplasma species (Mycoplasma orale, culturable and non-culturable strains of Mycoplasma hyorhinis) inhibited [3H]TdR incorporation and permitted the detection of some contaminated cell cultures that would otherwise have escaped detection in assays measuring [3H]TdR incorporation by mitogen-stimulated splenocytes. On the other hand, several other mycoplasma species (Mycoplasma arginini, Mycoplasma hominis) strongly enhanced [3H]TdR incorporation by unstimulated splenocytes. This enhancement was optimally detectable on day 2 after initiation of the cultures. The sensitivity of the assay was determined for a mycoplasma species (culturable M. hyorhinis) that inhibited as well as for one (M. arginini) that enhanced [3H]TdR incorporation. In both cases, the sensitivity was such that 1-3 x 10(2) mycoplasma colony-forming units (CFU) could be detected.
Alzani
R
R
Department of Immunology, Farmitalia Carlo Erba, Nerviano, Italy.
Rossi
R
R
Cozzi
E
E
Trizio
D
D
Marcucci
F
F
D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology / Q000382:microbiology; D002460:Cell Line; D003208:Concanavalin A; D005260:Female; D008213:Lymphocyte Activation; D051379:Mice; D008807:Mice, Inbred BALB C; D009174:Mycoplasma / Q000302:isolation & purification* / Q000502:physiology; D013154:Spleen / Q000276:immunology* / Q000378:metabolism / Q000382:microbiology; D013936:Thymidine / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D003208:Concanavalin A; D013936:Thymidine