Some of my cell cultures are contaminated, showing black spots that seem to multiply. The cells round up, and some of them appear to have granular membranes.
Any ideas what is causing this and how to eliminate it?
Hi
I think you face with mycoplasma contamination.
look at:
Article Prevention and Detection of Mycoplasma Contamination in Cell Culture
https://www.researchgate.net/post/How_can_I_decontaminate_mycoplasma10
https://www.bio-rad-antibodies.com/decontaminate-cell-culture-infection-prevent-contamination-mycoplasma-removal-agent.html
http://www.protocol-online.org/biology-forums-2/posts/26501.html
good luck
Hi Chris,
Alireza might be right about mycoplasma as an additional infection in your cells; however I would like to add some points to make it much more practical:
1- Mycoplasma is an intracellular infection and you will not see that small black moving things in your culture media.
2- If you are sure that they are moving, you might consider bacterial infection for your culture. If the bacterial infection is true, the source of infection might be the operator, the culture stuffs or whatever you use in this process.
3- I would suggest to put 5ml of your media in incubator for at least 24 hours to make sure that the medium is not the source of infection.
4- For rule out of mycoplasma infection, I would suggest Dapi staining.
5- In some labs they add p/s (penicillin and streptomycin to the medium to prevent infection).
6- In terms of treatment of infection, please make sure that infection or your treatment modality do not affect your readouts.
Hope it helps.
Regards,
Zahra
Thank you for the advice! I'll test the media as you suggested.
I'm already using P/S in my media, which I know is recommended against.
Looks like I need to scrap it all and start fresh. c'est la vie...
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