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Clumping of MCF7 cells - (Jun/15/2005 )

I'm having some trouble preventing MCF7 cells from clumping during passaging and single cell suspensions are necessary for further steps. I've tried trypsinizing for up to 10 min. at 37 degrees, pipetting up and down with a 1ml pipette, and even a bit of vortexing.
Help please!


What I do is, trypsinise the cells, put in 1-2 mL of their media (to neutralise it) and then use a syringe to de-clump. Run it throught a 20 (or 22) gauge needle 3-4 times.


after trypsinization, suspend the cells in media.
Put the suspension in a tube and wait for 1 min.
Use only the supernatant for passaging.


Donot tap the flasks after trypsinization.