I culture mouse stem cell on the feeders ,after ten days differention, I want to perform flow cytometry to check some markers in the cells, so I need to digest these cell into single cell, I know trypsle is much better to Trypsin,but I do not know how long I need to digest them?
does anybody can help me ?
It should not take much longer than the usual time for trypsin. We frequently use TrypLE express for detaching adherent cells and it works similarly to trypsin, within 5-10 minutes cells are detached after incubating with TrypLE at 37ºC.
You can check it with your own culture conditions by adding a proper amount to your cell culture and following the behavior of cells under the microscope until you see they have detached.
To help get single-cell suspensions you can also pass them through a 100 or 60 um cell strainer after detaching, and eventually use a syringe with needle for ressuspending into single-cells.
Cheers
In our lab we see that the incubation of Tryple for 5 to 10 mins is sufficient for disassociation of cells from plates.
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