Mycoplasma contamination impairs transfection efficiency

Contamination of cell cultures with Mycoplasma species can affect your cells in numerous ways, most of which will reduce the reliability, reproducibility, and consistency of your experimental results. Some of the general effects observed in eukaryotic cells can include:

Alterations to the levels of protein, RNA, and DNA synthesis due to nutrient deprivation
Changes in cellular morphology
An increase (or decrease) in cytokine expression

As a result of the various general effects on eukaryotic cells, data indicate that mycoplasma infected cell cultures have a decreased transgene expression compared to non-infected cell cultures.

The reduced transfection efficiency is most likely a consequence of several effects of mycoplasma on their host cells such as their lodging in the cell membranes and thus modulating cell membrane processes, or nutrient deprivation. Yin et al. demonstrated that the reduced transfection efficiency in mycoplasma contaminated cells is partly due to the deprivation of the essential amino acid L-arginine. This L-arginine deprivation is caused by the microbial enzyme arginine deiminase (ADI), an enzyme that is abundant in mycoplasmas. ADI is responsible for the catabolism of L-arginine by the mycoplasma.

The graph shows Hela cells transfected with GFP expression vector showing a significantly reduced GFP expression when cells are infected with M. fermantans.

To ensure your cells are free of mycoplasma, a regular biweekly test for mycoplasma is recommended. The MycoAlert^® Mycoplasma Detection Kit is easy-to-use and easy to integrate into a splitting routine. The assay provides results in less than 20 minutes by a simple 2-step assay. As a selective biochemical test, the assay exploits the activity of mycoplasmal enzymes which are found in all six of the main mycoplasma cell culture contaminants and the vast majority of 180 mycoplasma species but are not present in eukaryotic cells.

If you want to learn more about mycoplasma and their impact, we also have a webinar on mycoplasma contamination in cell culture.

Written by

Isa

Scientific Support Specialist

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