Wash cells or not after trypsinization? - (May/03/2005 )

I don't wash after trypsin-only using 2 ml in a 75 cm2 flask. After trypsinizing, I add 10 ml of media to flask which dilutes trypsin.

I think its always a good practice to wash your cells after trypsinizing. I never found something going wrong doing so. Additionally, when you spin down the cells during washing with PBS you get rid of dead cells and possible debris. It may be important to use PBS without Ca+2 and Mg+2 depending upon cell line.

I used to always wash after trypsination before, but stopped doing this, mostly to save time. Not been a problem so far. Just make sure the amount of trypsin you use is not to high (approx. 1:10 ratio of trypsin:medium). But when doing important experiments I still include the washing step!

i am not washing the cells after trypsinization and it workout for me

I only do the washing when I freeze cells. I know that is a short cut, but I can live with that.