I have to count live and dead cells after drug treatment. So I was wondering if resuspension (detachment) of cells with pipetting will affect cells or it should be first washed with PBS and then application of accutase (in this process I will lose cells because of washing).
Hello Kaneez Fatima
Gentle pipetting should be enough to detach HEK293T cells because with little agitation theses cells will detach from the substratum. Gentle pipetting will not damage the cells. Do not wash with PBS as you will lose cells in the process as you mentioned. If the cells detach easily then there is no need of using accutase.
Best.
I completely agree with Malcolm. I have also worked with HEK 293T and they are very loosely attached to the substrate, for this it is enough to pipette to detach them. However, avoid forming bubbles that could damage the cells and avoid pipetting very vigorously.
if you want an overview of the methods to use to detach cells, including pipetting and accutase, you can see here
https://cellculture2.altervista.org/subculture-of-adherent-cells/
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