I'm currently using amphotericin B, but it seems the fungus is resistant to this. Has anyone used nystatin, clotrimazole, or econazole? If yes, at what dosage? I'm using fibroblast cells.
Jhoti,
Unless your cell culture is irreplaceable the best thing to do is destroy the culture and purchase new cells. Eliminating Fungal contamination from a culture is nearly impossible. Plus handling the contaminated cultures just increases the odds of spreading the contamination not only to your clean cultures but other members of the lab culture.
Jhoti,
Unless your cell culture is irreplaceable the best thing to do is destroy the culture and purchase new cells. Eliminating Fungal contamination from a culture is nearly impossible. Plus handling the contaminated cultures just increases the odds of spreading the contamination not only to your clean cultures but other members of the lab culture.
Contaminating cultures - especially if they are from rare cell populations are a problem. I agree with Daniel Medina that if you can remove the culture and start again this is the best - avoid cross contamination of other cultures - and if the fungus is aggressive, can sometimes even lead to the entire incubator becoming contaminated long term.
However, my favorite agent is Primocin: http://www.invivogen.com/primocin
It is very very good for bringing back precious cultures contaminated - especially during any long term stable cell line selection.
Good luck.
Hi, Jhoti, I agree that the best way is to get new culture. Howevre, you can try also several presverative mixture: http://www.plantcelltechnology.com/about-ppm/
Of course, you can try nystatin and other antibiotics, but antibiotics may have effect of fibroblast cells as well. Good luck!
hi Jhoti, i strongly agree with all the above answers. The best thing you can do with fungal contamination is to discard the culture, properly sterilize the culture room, culture vials and all the utensils using for culture and fumigate both the incubator and culture room.
If your culture is a are one and you have no vials to spare, you can subject your culture flask with 4-5 time wash with PBS containing higher doses of a mixture of anti-mycotic agents. Then replace the PBS with media with lower doses of the same mixture. Repeat the steps for a couple of days until you get your culture free of fungus.
Just discard the culture and start fresh. No amount of treatment will remove the contamination and keeping bad cultures means you run a huge risk of contaminating any other cultures keep in the same conditions. Do you add an antibacterial/antimycotic to your culture medium for your FB?
Dear Anne, Yes i add usually add strept-penicillin and amphotericin B to my medium, should i stop using this during routine culturing? May this have cause some kind of resistance?
We regularly culture primary human skin cells in our lab (MC, KC and FB). We add 5ml of PSF to 500 ml DMEM for FB cultures. The PSF we purchase is from Corning (30-200-CI). You need to continue adding this to all your culture media otherwise it will be more likely to become contaminated.
Dear Anne, Would you give me full name of PSF, I can not find Corning (30-200-CI)
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