Bacterial Contamination in HuEmbryonic cell lines seen in SFM4 but not in DMEM +SFM4 DMEM + 10% FBSm Bacterial Contamination
Posted 02 January 2013 - 08:00 AM
I have transfected HuEmbryonic cell lines going. They appear to do fine in DMEM + 10% FBS media but as soon as I change the media in 70 - 95% confluent cell lines to SFM4 (commercially available), The cells die due to bacterial contamination.
I see this trend in almost all my transfected cell lines. I do not use any antibiotics. But the DMEM plates are healthy and show no trace of contamination even when passaged. And this is not an isolated incident, it happened 4 times already viz. as soon as media is changed bacteria crops up.
I have done bacterial assays against the SFM4 media control, cells in DMEM + 10% FBS and the contaminated cells in SFM4. Only the contaminated show high readings everyhting else is negative for bacterial contamination.
Any help or suggestions would be greatly appreciated.
Thanking you all in advance.
Posted 02 January 2013 - 02:08 PM
Posted 03 January 2013 - 02:15 PM
You should also go through the incubators and give them a good clean, change your lab coat, throw out any opened stocks of medium, PBS, trypsin, and other reagents you are currently using. Appraisal of your sterile technique would be a good thing to do as well, especially if you are new to cell culture.