Dear All,
What is the ratio of FBS to trypsin to inactivate/neurtrilize trypsinization in cell culture?
In some protocol, the ratio of 10%FBS media to 0.25%trypsin-EDTA is 1:1 or 2:1. ATCC guide uses 3:1. So what is minimum or the best ratio of FBS to inactivate trypsin.
Thank you very much
Cheerio,
I think the best ration under this condition to inactivate trypsin is 7: 1.
What concentration of trypsin do you use? I generally use 4:1 or 5:1 medium to trypsin whe using 0.25% trypsin. I also always centrifuge, aspirate the medium containing trypsin and plate in fresh medium. This step is very important, because any active trypsin remaining will irritate your cells. If you do not centrifuge and remove the trypsin containing medium, then you might consider neutralizing the trypsin with a small amount of pure FBS before adding medium and re-plating to be sure that you have successfully neutralized all the trypsin.
To my experience, I use 1:1 ratio and centrifuge. After I remove the supernatant that contains trypsin and media, I resuspend with new media or PBS and go from there. If there is any trypsin in the resuspended cells, it's no more than 100ppm.
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