Hello Everyone,
We are facing a problem in all cell lines running in our culture room. After two or three passages, the cells start showing the granules like structure on its surface. They propagate properly, and there is no turbidity in the media as well. But after some more passages the background debris become very prominent and cells start dying. Can anybody suggest, what is the problem. We have changed, almost everything and also fumigated the culture room. I am attaching the picture of the cells at the third passage.
The problem is persisting with all the cells growing in the lab HEK-293, SH-SY5Y, HT-22 and CTC12. Media is DMEM high glucose with 10% FBS and anti-anti is common in all cells media.
Antibiotics could mask the contamination, which still can affect your cells. I would recommend to subculture the cells to antibiotics-free media, and if contamination appears, you can say it was caused by contamination. If no contamination occurs after culturing in atb-free media, you are almost sure it is not caused by (regular) contamination. In that case, the particles found in media could be just fragments of dead cells caused by e.g. wrong cultivation conditions.
BTW, if you can, try to avoid using antibiotics routinely.
I have no experience with viral infections (It might be the case?).
Have luck.
Jakub
Hi Ibrar,
Have you investigated a potential problem with mycoplasma?
Stain your cell with Dapi.If you can see little blue dots, that means you have mycoplasma contamination.
Good luck!
Alice
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