Which trypsin EDTA to use? - (Jun/21/2010 )
I've been culturing some human granulosa cells for a while now and I normally use 0.05% trypsin 0.02% EDTA in hanks balanced salt solution with phenol red to trypsinise them. Last time I trypsinised them I had run out of this trypsin so I used 10x 0.5%trypsin 0.2% EDTA diluted to 1X with PBS instead thinking it would have the same effect. Only a few of the cells would detatch and they ended up clumping together so I had to throw all of the cells away! Does anyone know why the cells reacted differently to this trypsin when the concentrations were the same?
punkyneen on Jun 21 2010, 04:41 AM said:
was the source of the pbs clean? i.e. no lps?
Does your PBS have magnesium and calcium? Mg and Ca promote cell adhesion.
In addition to navjay's comment; EDTA captures Mg and Ca, so the result is probably that your cells detach from the plate but clump together.