Macrophage again~~ Please help~! - (Sep/25/2007 )

Hi, all.
I've got macrophages out from peritonium without injecting Thio.
The thing is..They DETACH after 24hrs [They adhere well after 2hr incubation, but I'm not sure when they become detached]. Is this normal????
I use RPMI+10%FBS+1%pen/strep+1%pyrubate+2-ME.
Please help~!!
Thanks a lot~~!!

i had a similar problem - i found that lowering serum concentration helps a lot and you should also check if the nun-adhering cells are indeed macrophage (i think staining them with CD11b is the easiest way).