Yeast in Cell Culture... - How to erase the Yeast? (Jun/06/2002 )

One of my cell cultue is contaminated with yeasts.
Does anyone know a way to eliminate them from culture without harming my fibroblasts? Usually I would throw them away, but since these are my only cells of this kind and i really need them I would linke to save them!
Thanks in advance!

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Hi Kathrin,
I had the same problem with primary astrocytes cultures.

What you can do as last resort is to wash 3X with PBS, then incubate your cells with Nystatin or Fungizone (Amphotericin B ) and normally you should see the difference appearing by stopping the yeast growth.....

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hi
here is a protocol described in this forum. I've no tested it myself, but i was reported it works...

Decontamination of cells from the yeast
I Destroy yeast
1. Aspirate medium and wash cell in PBS.
2. Incubate cells at 37oC for 5 min in non-diluted antibiotic-antimycotic.
3. Incubate cells at 37oC for 5 min in trypsin. Spin them down (100g, 5 min). DO NOT EXCEED 100g.

II Secure cells from crosscontamination
4. Slowly and carefully aspirate supernatant. Resuspend cells in 0.5 ml of regular medium.
NB! We assume that your regular medium contains 1x antibiotic.
5 Transfer cells to a new tube and dilute them with regular medium to 6 ml.

III Prepare cell cultures for regular culturing
6. Seed cells on 30 mm Petri dishes: 1:1, 1:3, 1:7, 1:15, 1:31, 1:63 and 1:127 (cell suspension/regular medium).
7. Change the medium before you leave for home.
8. Next day: check for results

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