Mycoplasma Contamination in Cell Cultures
One of the most common contaminants present in
cell culture laboratories are mycoplasma. A conservative estimate states that
between 15-35% of all continuous cell cultures are contaminated with mycoplasma1,
some estimates are even higher (up to 80% in some countries2). Read more...
Effects on Your Cell Culture
Typical routes of infection are cross-contamination from
untested infected cells (e.g. via aerosols generated during pipetting, use of
same media bottles, handling of more than one cell type at one time),
contaminated materials, contaminated donor tissue (<1%) or direct infection
from the researcher. The primary source is normally cross-contamination from
infected cultures. Mycoplasma grow slowly and do not kill the cells outright
but affect various cellular parameters 1,3,4,5,6,7,8,9,10,11 (see
figure). Thus, mycoplasma contaminations can seriously impact the reliability,
reproducibility, and consistency of experimental results, representing a major
problem for basic research as well as for the manufacturing of bioproducts.
Standard testing for mycoplasma is an important quality control.
Mycoplasma – The Invisible Enemy
Even at very high concentrations (> 107
cfu/ml) mycoplasma are not visible by microscopy. They do not cause visible
changes in the growth media that are commonly associated with bacterial and
fungal contamination, such a turbidity or pH changes1.
Therefore contaminations are very difficult to detect and the presence of
mycoplasma can remain undiscovered for months.
As mycoplasma compete with cells for the nutrients in
culture media, one of the first visible signs is a slowdown in cell
proliferation. Other indications of contamination include cell aggregation,
morphological changes or poor transfection efficiencies with cells that
originally transfected well.
Eliminate and Prevent Mycoplasma Contamination
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1) Drexler HG, Uphof CC (2002). Cytotechnology 39: 75–90
2) Koshimizu K, Kotani H (1981). In: Procedures for the
Isolation and Identification of Human, Animal and Plant Mycoplasmas (Nakamura,
M., ed.), Saikon, Tokyo, 87-102
3) Gong H et al. (1999). Biochem Biophys Res Comm 261:
Ben-Menachem G et al. (2001).
FEMS Microbiol Letters 201: 157-162
5) McGarrity MF et al. (1984). In Vitro 20: 1-18
6) Sokolova IA et al. (1998). Immunol Cell Biol 76:
7) Doersen CJ, Stanbridge EJ (1981). Mol Cell Biol 1:
8) Stanbridge EJ (1971). Bacteriological Reviews 35):
9) Darin N et al. (2003). BMC Biochem 4:15
10) Rottem S (2003). Physiol Rev 83: 417-432
11) Miller C et al.(2003). Biotechniques 35:812-814