cell clumps!!

Brett Lindenbach brett at BORCIM.WUSTL.EDU
Wed Dec 2 12:39:47 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.
>>>
>>> thanks
>>
>>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
>>cell clumps.
>>
>>Claire
>>
>
>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
>longer.
>
>        - Dima
>
Actually, the best way I have found is to buy some of those sterile disposable
plastic sieves that fit perfectly into the top of a 50 ml Falcon conical. They
will retain large clumps and be broken up by showering the residue with media
to release additional single cells.  Dima's plan sounds too barabaric to me.
;-)

Brett Lindenbach
brett at borcim.wustl.edu
Dept of Molecular Microbiology
Washington University School of Medicine





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