Tissue Culture Core Facility (TCCF)

Services

Quality Control of Cell Culture with Mycoplasma Testing

Studies have repeatedly shown that the incidence of mycoplasma contamination ranges from 25% to 87% of cell cultures (G.J. McGarrity and H. Kotani. In: The Mycoplasmas, Vol. IV. S. Razin and M.F. Barile (eds) Academic Press. pp. 353-390, 1985). Mycoplasma can be introduced into cell cultures by laboratory personnel and into newly introduced contaminated cell lines as well by reagents. Their small size (approximately 0.2 µm) allows them to pass through most 0.22-µm sterilization filters, and they are resistant to common antibiotics such as penicillin and streptomycin (C. Lincoln and M. Gabridge: Methods Cell Biol. 57:49-65, 1998).

The TCCF is recognized as being vigilant in monitoring cell culture quality, giving advice to investigators on handling and, importantly, preventing mycoplasma contamination. Investigators are encouraged to use antibiotic-free media to
prevent the development of antibiotic-resistant bacterial and mycoplasma strains and to avoid the effects of antibiotics on cellular metabolism.

MycoplasmaContamination by mycoplasma species can easily lead to unreliable and unreproducible results/experiments (J Tang et al. Microbilogy Methods 39: 121-126, 2000).

MycoplasmaSeveral methods have been developed to detect mycoplasma contamination.

The TCCF uses nested PCR and fluorescent microscopy simultaneously to ensure the best and most accurate results. The primer set used allows detection of various Mycoplasma species (M. fermentans, M. hyorhinis, M. arginini, M. orale, M. salivarium, M. hominis, M. pulmonis, M. arthritidis, M. bovis, M. pneumoniae, M. pirum and M. caprocolum), as well as Acholeplasma and Spiroplasma species, with high sensitivity and specificity (an example is shown in the figure below). TCCF also offers mycoplasma testing of culture media and serum and media supplements.

   

Questions? Please contact Steve Boldogh, Director at (409) 772-9414 or Yixiao Sun at (409) 772-4903.