I need to do cell proliferation assay for LNCaP cells after stimulation with androgen. The protocol I follow is I grow the cells in 96 well plate in 10% complete medium for 48 hrs. then remove the medium, give a PBS rinse of the plate and then add medium with 2% charcoal stripped serum. After adding medium I stimulate with different doses of androgen. However, the problem is, since LNCaP cells are loosely adherent cells, they tend to come out when I aspirate medium or rinse with PBS. They also come out when I add fresh medium with 2% CS FBS. As a result, the cell number varies.. which in turn will give me a wrong cell proliferation index. IF somebody cud help me on this topic..
Edited by sheebs, 07 January 2010 - 12:41 AM.