klenchin at REMOVE_TO_REPLY.facstaff.wisc.edu
Tue Dec 1 19:47:57 EST 1998
In article <366400DA.759D048 at sghms.ac.uk>, Claire Fenech <c.fenech at sghms.ac.uk> wrote:
>> Hi all,
>> i was wondering if anyone can get rid of cell clumping together after
>> trypsining them of the tissue culture dish?
>> I normally can get them of the tissue culture dish no problems but the
>> cells still sticks to gether making them hard to count. is there a good
>> way to get over this problem.
>If your cells are not too fragile you can resuspend them in fresh medium
>(small amount 1,2 ml) and "triturate" them gently: sucking them up and down
>a sterile, fire polished glass pasteur pipette. Keep the tip of the pipette
>close to the bottom while you draw or expel the cells. This dissociate the
And for even more stringent trituration which virtually guarantees that over
90% cells are single, you can jam sterile yellow tip on the 5-10 ml pipet
and do ~ 10 passes. Works great (kills some % of cells though).
Other suggestions also apply - split before confluent and trypsinize
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