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[punkyneen]

Hi everyone,

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?

Thanks

[telamon]

punkyneen, on Jun 21 2010, 04:41 AM, said:

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...
Thanks

was the source of the pbs clean? i.e. no lps?