Following differentiation experiment (24h PMA 100 ng/mL -> 48h RPMI 10% FBS -> 72h IL-4 40ng/mL (M2) or LPS 50 ng/mL + INF 50 ng/mL (M1)) we found very difficult to detach cells for FACS analysis of CD163, CD209 and CD80. We used 5mM EDTA at 37oC for more than 20 min. Only half of cells detached. Can Trypsin be used in this situation? Any other advices?
Tran Canh Tung Nguyen
Hello Honorata Kraśkiewicz,
Activated THP-1 cells are highly adherent, and using Trypsin may also hamper cell surface markers. So, I am using Accutase and incubate at 37˚C for 30 mins. This works quite well. It can help you get about 80% of the cell.
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