After fumigation of co 2 incubator, I thawed the cell line from where I used to thaw before too and after 48 hours media color changed to yellow and turbid. I used rpmi media from another lab and they growing easily cells with the same media. Any suggestion would be appreciated. Plz suggest.
sometime it takes more time to adhere, or you can increase serum concentration. but increasing serum can also increase the chance of contamination.
Hi, I think first you should check with just media in incubator if it looks fine then may be cells got contamination during frozen so you may try to get cells from other source and culture.
Hello Arpita Dey
I agree with Mamatha Serasanambati . Are you sure the CO2 incubator is free of contamination? You can keep agar plates with open lid in the incubator and check after 24 hours for contamination.
Additionally, CO2 incubator must be cleaned with 70% ethanol regularly. The water tray present in the CO2 incubator should contain a disinfectant/detergent in water to prevent bacterial growth.
We use DMEM media to grow Mcf-7. you can use MEM also. As it has slow growth, it takes time to get confluent (about 4days). During thawing, you can use 20% FBS in media for speedy growth. If less contamination comes, you can see the media clean but dotted particles in the flask under microscope. For that you can remove the media and wash the flask with trypsin+media (1:3) and again add media and keep it in incubator for 24hrs. If still contamination comes try to passage again. But if the media become fully turbit again, discard it.
Arpita Dey media getting turbid and yellow is also a sign of fungal (yeast mostly) contamination. for MCF-7 you should use DMEM with 10% FBS. This cell line is rapid growing so may be you have inoculated too many cells and that's why in 2 days your flask got overconfluent and cell started floating and dying ( also explains why media got turbid and yellow)
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