Hello Zarin Sharifnia!

Are you using treated flask, a way to reduce adhesion is use non treated petri dish/flask. Or you can try to use accutase to remove the cells, this is a reagent less agressive than trypsin.

Good luck with your experiments

Zarin, another method to try is to use ice-cold PBS for a few minutes to help remove the cells.  Pour/pipette off the culture medium, rinse with PBS to remove any remaining medium, and then incubate for a few minutes with ice-cold PBS.  Sometimes to the cold and serum removal can get them to let go a bit easier.  When I was working frequently with macrophages (which are also very adherent), I also cultured them on non-tissue culture treated bacterial plates and they were easier to remove from there.  For some experiments where their surface markers were integral to the outcome, I also bought some special NUNC culture plates that were adherent at 37 deg, but non-adherent at room temp and below.  They're more expensive than normal plates, but they did work.

One possible answer is that when you add IL4 or GM-CSF you may be modifying pH. Check pH before and after adding IL4 and GM-CSF. If there is a notorious difference I suggest you introduce a buffer in the media to stabilize pH.

Hello Dear Friends

Thank you very much for your suggestion.So far I had not thought to change PH.I must try.