I usually filter sterilise the media w/ FBS or FBS alone (post heat inactivation). I use 0.22um PVDA membrane filters or syringe filters at times. I also use sensitive cell lines. The cells lines are gathering more dead cell debris. It made me wonder, whether the filter sterilisation step is devoiding the media of certain FBS constituents.
Normally, depletion of components, if any, is minimal to cause any problems in most cell lines. However, if you suspect that this might be the case, test it by filtering the medium without adding the FBS and then add the required amount of FBS which is always supplied as sterile solution.
In my lab, we never filter sterilize the medium with FBS. If your sterile technique and the setup is decent, filter sterilization of working dilutions of medium is an overkill. In many cases, we do not use Pen/Strep in the medium.
We don't filter our FBS after heat inactivation. If you are worried about the precipitates, you can centrifuge FBS for 5 min to get rid of them.
Hello Gargi.
We always filter sterilize our FBS before we use it. And we never had any problems till now.
Do you see the debris after thawing the cells? If so, normally the debris disappears after the first wash and passage. But maybe you should check your cryoconcervation- and revitalization steps.
Greetings
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