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Differentiated THP-1/U937cell removal from tissue culture plate | lak001 | 2010-05-05 | Click here to register. |
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I've been trying to remove differentiated THP-1 and U937 cells from tissue culture plates with some difficulty.
I stimulate the THP-1 with 100nM PMA for 3 days and then let them rest for two days before trying to remove. The U937 cells are stmulated with 100nM for two days and rested for 3 days before trying to remove from the plates. They are tissue culture treated plates. I have used 5mM EDTA and Cellstripper (Mediatech, non-enzymatic).
I pipette up and down after incubation with the disassociation reagents and the cells are very hard to get off the plate. I get about 60-70% vivability of the cells that do come off too, although I have not checked the viability before trying to remove. The cells look healthy and are obviously metabolizing since the medium changes color pretty quickly (overnight). I don't want to use enzymatic disassociation reagents because I need to use them in functional assays immediately after removal from the plates. Has anybody had this issue and have a remedy for it? I don't want to use a cell scraper because I feel it will just damaged the cells more. |
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