I'm trying to evaluate several cytotoxicity endpoints upon internalization of silica particles on macrophages differentiated from THP-1 cells, but we are having some issues with the detachment of cells from well, during the harvesting phase. We currently use a solution of EDTA with trypsin in PBS, but results are not satisfactory.
Trypsin is something you cannot use with THP-1 cells. Use PBS with 1mM to 5mM EDTA to lift differentiated, adherent THP-1 cells from plates. Some protocols have used Accutase for this purpose. But, PBS/EDTA works pretty well.
This is what we used to do to detach macrophages from a plate: Add 10 mL pre-warmed PBS and leave in the incubator for 10 minutes. Tap the plate firmly and this should detach most cells. Gently scrape off the rest (if needed). If this strategy fails, try accutase. I have heard from others that it works well.
Sanath is right. Cold PBS-EDTA works pretty well. Ready to use solution Versene is my daily tool for that!
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