Can I use Nystatin (100 U/ml in final concentration) or Amphotericin B for treatment of yeast contaminated cell culture plates to maintain the cells for analysis within 10 days after contamination or discard them.
It is always preferred to discard contaminated culture, improve cell culture techniques and start freshly. However, it is not easy to discard a culture plate half way through an experiment OR when repeatedly gets contaminations.
It is more likely to get contamination when plates are taken out frequently for analysis or left outside for prolonged time (e.g. life cell imaging). It could be either bacterial or yeast contamination. You specifically mentioned yeast contamination and I presume you are sure about the organism. I have treated cultures with "fungizone" without any major adverse outcomes. Some add a fungicide when establishing cultures from skin biopsies as a prophylaxis. For long term cultures, it is important to adhere to protocol and change medium with fresh fungicide at recommended concentration. If it is a short term culture (and these cells will not be recovered for other purposes), developing resistance would not be an issue. I hope that if you will find out the half life of the fungicide and add it appropriately in between. Please also make sure that intended fungicide will not interfere with outcome measurements. Good luck.
It is always preferred to discard contaminated culture, improve cell culture techniques and start freshly. However, it is not easy to discard a culture plate half way through an experiment OR when repeatedly gets contaminations.
It is more likely to get contamination when plates are taken out frequently for analysis or left outside for prolonged time (e.g. life cell imaging). It could be either bacterial or yeast contamination. You specifically mentioned yeast contamination and I presume you are sure about the organism. I have treated cultures with "fungizone" without any major adverse outcomes. Some add a fungicide when establishing cultures from skin biopsies as a prophylaxis. For long term cultures, it is important to adhere to protocol and change medium with fresh fungicide at recommended concentration. If it is a short term culture (and these cells will not be recovered for other purposes), developing resistance would not be an issue. I hope that if you will find out the half life of the fungicide and add it appropriately in between. Please also make sure that intended fungicide will not interfere with outcome measurements. Good luck.
you might be able to put in a mixture of chitinase, beta-glucanase and chitosanase, completely non-toxic to eucaryotes, but should lyse all yeast and fungi.
It would be best to start fresh. The yeast contamination and the addition of fungicides will have an effect on the cells. The assay outcome for contaminated cells will more than likely be different than the outcome from cultures that are not contaminated. If using contaminated cells, how will you know if the result is from the experimental conditions or a result of the contamination? How could you be sure that the contamination had no effect on the results? When repeating the experiment to confirm the results, would you have to intentionally contaminate the cells with the same yeast in order to reproduce the results? A fresh start after decontaminating the incubator will give you more confidence in the results and increase the chances that you can reproduce the results.
Yeast-contaminated cell culture should be discarded immediately, and start your cell culture fresh from your cryo-stock
Candida? Destroy...
ALWAYS streptomycin +Penicillin G (Sodium) + Amphotericin B
Ratneswary Sutharsan gave you good and certainly more 'soft' answer.
Good luck.
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