Re: adherent cells

Al Sabirsh (
Fri, 13 Jun 1997 13:27:40 +0000

> Could anyone suggest a method of removing adherant cell lines from culture
> vessels which does not remove surface adhesion molecules? At present i am
> studying surface cell adhesion molecules expression on human umbilical vein
> endothelial cells using flow cytometry.
> i would appreciate any suggestions.
> thanks
> Morgan

Sure - we use adherent cells all the time, including HUVEC. Remove the
culture medium, rinse with a volume of PBS (that does not contain
calcium or magnesium) equal to your culture medium volume. Then
incubate the cells with a sterile filtered PBS solution containing 2mM
EDTA (this should be added at about one tenth of your culture volume).
Incubate for 10 minutes or so - you will probably have to jar the cells
loose from the plates. This is not as fast or effective as trypsin for
dissociating cells, but it will preserve surface receptors. (The cells
dissociate because they require the divalent cations, that are removed
by EDTA, for adhesion). You can then dilute the cells in PBS or cell
medium depending on what you would like to do with them. If is often
helpful to pipette them up and down a few times during this step to
disperse clumps of cells and to make sure that they are completely

Sigma also sells something they call "non-enzymatic cell dissociation
solution" (which I suspect is exactly the same thing as I described
above) if you're inclined to buy that instead.


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