Hi all,
Recently I encountered some irregular black/grey patches inside my cell culture flask (see attached image, x400 magnification). They are not motile, not floating, do not stick to cells but to the flask surface, cannot be removed by washing / trypsinization. My culture media color is always clear, not turbid and pink (not acidic).
Do you think it is contamination of sign of cellular stress?
Hi! It could be contamination with mycoplasma, because this type of bacteria doesn't generate turbidity and the culture media color is clear. Mycoplasma is difficult to identify under the mycroscope due to their small size, what you can find is an alteration of the cell morphology due to the contamination. Anyway, to be sure that you have mycoplasma in your culture you can do some tests like ELISA, PCR, microbiological culture or use some kits specialized for mycoplasma detection like PlasmoTest from InvivoGen, MycoAlertTM PLUS Mycoplasma Detection Kit, or MycoFluor™ Mycoplasma Detection Kit from ThermoFisher, and follow the protocol.
Here are the links where you can get more information about the kits:
https://www.invivogen.com/plasmotest
https://www.thermofisher.com/order/catalog/product/M7006#/M7006
Also here's some information about cell culture contamination I hope it can help you
https://safety.fsu.edu/safety_manual/supporting_docs/Understanding%20and%20Managing%20Cell%20Culture%20Contamination.pdf
Best of lucks!
Hi Ariadna Hernández , thanks for the answer. I have discarded these cells and thawed new batch of cells. Now everything is back to normal and hopefully it will continue to be normal =)
In my opinion, It seems to be some kind of fungi. Some of them just appear as tiny dots or colloids in the media, grow very slowly and they are very hard to remove. The best choice is to take a new sample as you did.
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