- I need to inoculate my cells in rat. However the culture has some bacterial contamination
- We use some antibiotic, however at beguin has effect, but now we observed some bacterial growth again
What can I do
Thanks
Burn all the cells and media and go back to a stock of cells you know are clean.
Trying to recover a cell line that is contaminated is not a good use of time and resources from what I have heard: you'll never know if the contamination is going to come back even if you find an antibiotic that clears it up. It might have a mycoplasma contamination which would invalidate your results depending on what you're doing with them.
I have been told that it's best to use NO antibiotics in cell culture so that if you do have a contamination, you know immediately and can purge the contaminated cells and will not have to guess what is contaminated and what is not. So far it is working out for me. I had one contaminated well since I stopped using antibiotics and have confidence none of my other cells are contaminated.
Antibiotic-Antimycotic (100X) - Thermo Fisher Scientific
First of all, Which type of antibiotic are you using? It is not recommended to use that batch of cells. If the infection is persistent you probably need to use another batch and discard the infected one. If you don't have another batch you can try with
What antibiotic are you using?
Do you filter sterilize your media before you use it?
If you see contamination, it is better to just throw out that batch and re-thaw a new vial of cells.
You should keep your culture rom bacer- free, for examole, open UV light for at least 20min before start. OPERATION STRICTLY ACCORDING TO PROTOCOL. Everthing item shold be new from now. OLd CULTURE AND CELL LINES YOU STORD, GIVE UP
Burn all the cells and media and go back to a stock of cells you know are clean.
Trying to recover a cell line that is contaminated is not a good use of time and resources from what I have heard: you'll never know if the contamination is going to come back even if you find an antibiotic that clears it up. It might have a mycoplasma contamination which would invalidate your results depending on what you're doing with them.
I have been told that it's best to use NO antibiotics in cell culture so that if you do have a contamination, you know immediately and can purge the contaminated cells and will not have to guess what is contaminated and what is not. So far it is working out for me. I had one contaminated well since I stopped using antibiotics and have confidence none of my other cells are contaminated.
Hi, Do you filter sterilize your media before you use it?
I wish to suggest that if possible use selective medium to grow your culture. Many selective media have antibiotics.
You may also identify the contamination and add antibiotics specific for that bacteria.
The underlying presumption is that all this is being done under aseptic conditions.
The best option is to discard your cells after adding bleach to your medium and to thaw a new cell sample.
Just before doing that, try to desinfect your incubator by ethanol 70%.This is might eliminate most of bacteria and avoid any eventual contamination.
Maybe there are spores of bacteria after that you culture your cells so you can eradicate them.
+1 to all the start over suggestions. In the future you could use commercially available antibiotics like PennStep to reduce the risk of infection. Also, the contamination you have might be yeast or another fungus, if you're already using PennStrep. I recently switched from PennStrepp to Anti-Anti (contains an anti-fungal) to reduce the risk of yeast contamination in my cultures.
Antibiotics and antimycotics won't solve your problem, and they won't gurantee it will never happen again, but it's a good safety measure to reduce the risk of contamination (barring very, very bad technique).
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