Mycoplasma are classified as prokaryotes with several unique properties that distinguish them from other prokaryotes. They lack a cell wall, instead using sterols, especially cholesterol, from vertebrate hosts to maintain their plasma membrane. Therefore, they are unaffected by antibiotics that interfere with the murein formation of cell walls. It also means that mycoplasma do not overgrow their host cells but in fact bind with cell walls to obtain nutrients. As they are extremely small (0.15–0.3 μm), these organisms are difficult to filter out of suspension, and can grow to particularly high concentrations in mammalian cell cultures without producing turbidity or other obvious symptoms.
Mycoplasma grow very slowly as they infect the cell culture. Nonetheless, this can have a serious effect on the host cell’s metabolism. Unexpected alterations can develop in growth, metabolism, function, secretion and synthesis, and expression of the host cell. Additionally, damage can occur to the host cell’s membranes, DNA and RNA, and other intracellular organelles. All of this can lead to skewed and/or inaccurate data and, if not recognized as a mycoplasma infection, can seriously compromise the validity of the final product or study results.
The only thing you need to do is bring 1 ml of supernatant to the BioNanoLab preferably in a 1,5 ml tube (epje).
This can be:
1. Cell supernatant during passage of suspension cell culture.
2. Supernatant from adherent cells prior to trypsinization.
3. Cell supernatant from cells brought “up from frozen” – cells out of liquid nitrogen with uptake into media. Leave minimum of 1-2hr before testing under normal culture conditions.
NOTE: Cells diluted into fresh media after passaging or trypsinization result in a much lower signal - leave minimum 24hr under normal culture conditions before testing. Supernatant can be stored at room temperature or 4°C for testing the same day.
Supernatants can be stored at 4°C for ≤ 5 days but do not freeze supernatant.
The first test is planned for April 26th.
You can bring the samples on Monday April 25th.
Results can be expected by email within a few days.
The costs per sample will be 15 euro.
Please inform Irene Konings (firstname.lastname@example.org; phone 3593; office ZH151) if you are interested in joining the mycoplasma test, and how many samples you want to bring?