Getting cells off plates

Tom Vink tvink at xs4all.nl
Fri Mar 29 00:33:23 EST 1996


ez022056 at dale.ucdavis.edu (Edward Wang) wrote:

>Pearse Ward (ward0101 at tc.umn.edu) wrote:


>: I'm having a problem getting COS-7 cells to dissociate from plastic 
>: tissue culture plates.
>: 
>: I am doing expression cloning and selecting transfectants by FACS. The 
>: antigen I am detecting is very sensitive to trypsin, so that is not an 
>: option.
>: 
>: I have tried Versene, and PIPES/NaCl/EDTA, and both work to a degree 
>: with incubation times of 15-30 minutes, but neither one removes all of 
>: the cells from the plastic.
>: 
>: I'd be interested in other people's favourite recipies for non-enzymatic 
>: dissociation of cells.
>: 
>: Many thanks,
>: 
>: Pearse
>Couldn't you just scrape it with a cell scraper?  Fisher sells them for 
>dirt cheap, and that's what I've always used to take cells off plates 
>(trypsin usually makes a mess...
>-- 
>********************************
>Edward H. Wang                 *
>Staff Resch. Assc.             *
>Dept. Internal Med. -Nephrology*
>UC Davis                       *
>********************************
Trie using 100 mM EDTA (probably too much) in water, instead of a
buffer this did wonders for us when removing endothelial cells instead
of PBS/EDTA
Tom Vink
Department of Haematology
University Hospital Utrecht
The Netherlands




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