Dear all,
Regretfully, I am exhausted and angry about bacterial contamination of my sweetheart cell lines. I have checked and changed anything including Pasteur pipette, pipette tips, culture media, incubator, and even freezed cells. But I could not find what is the sources of contamination.I hate disgusting bacteria! Dose anyone know how can I get rid of these insidious and harmful microorganisms. Most of bacteria are Gram+. Thanks in advance for your comments and helpful hints.
Kind wishes
Alireza
Dear Alireza,
Do you have the same problem with other cells in that culture room as well or specific to the particular cell line?
You changed everything except the cell culture hood based on the information provided by you. And sometimes, you need to sterilize entire cell culture room. And also, the cell culture hoods can be home for insects, which you can not see. I would recommend cleaning your hood properly including the SEPA filters.
However, I would strongly suggest fumigation of entire cell culture room while you left the hood running and the incubators open during the process of fumigation.
The cheaper way to do fumigation is by using fumigation mixture 35 mL of formalin (40 percent formaldehyde) plus 10 g potassium permanganate per cubic meter of space. You need to seal the room completely and protect your reagents properly before fumigation. You can not use the room for three days. After three days you need to ventilate it for few hours before using the room.
Please find more information in the following link for more methods and details.
http://www.fao.org/docrep/005/AC802E/ac802e0j.htm
I hope this information is helpful to you. Good luck.
Dear Alireza,
Do you have the same problem with other cells in that culture room as well or specific to the particular cell line?
You changed everything except the cell culture hood based on the information provided by you. And sometimes, you need to sterilize entire cell culture room. And also, the cell culture hoods can be home for insects, which you can not see. I would recommend cleaning your hood properly including the SEPA filters.
However, I would strongly suggest fumigation of entire cell culture room while you left the hood running and the incubators open during the process of fumigation.
The cheaper way to do fumigation is by using fumigation mixture 35 mL of formalin (40 percent formaldehyde) plus 10 g potassium permanganate per cubic meter of space. You need to seal the room completely and protect your reagents properly before fumigation. You can not use the room for three days. After three days you need to ventilate it for few hours before using the room.
Please find more information in the following link for more methods and details.
http://www.fao.org/docrep/005/AC802E/ac802e0j.htm
I hope this information is helpful to you. Good luck.
Dear Ramachandramouli,
many thanks or your useful information. But, I should add this point that our laminar flow II cabinet is shred by other technicians and fortunately their cells are happy and free of any bacterial contamination. However, your illustrations regarding fumigation was so perfect and informative. Hope all the best for your Post Doc in Hannover:)
Kind Wishes
Alireza
I assume that the problems come from either your handling process or the frozen cells themselves.
I would suggest you take some cells from the technicians you have mentioned and maintain by yourself and also give your frozen cells to them and ask them to maintain. Then, you will come to know about the problem exactly. If the cells contain bacteria already, I would suggest you go for including additional antibiotics in your culture medium or get a fresh stock from others.
Goodluck
- Badges
- Science topic
- Similar topics
- Biological Science
- Microbiology
- Microorganisms