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macrophages subculture - how can i subculture monocytes/macrophages (Jun/17/2006 )

hello everybody!!!
maybe somebody could help. I started to work with PBMC derived monocytes/macrophages. How can i remove the macrophages from the culture dishes if i want to continue subculture them. I tried long Trypsin-EDTA treatment with following extensive up and down pipetation, it's doesn't help. Cells were attached very strong to the dish.

if i would scrab them with a scraber, does it could damage the cells?

thank you,

-person B-

hi person B ! method of subculturing depends on the cell type. macrophages adhere very stongly to plastic surfaces. infact it is this property of macrophages which allows us to separate them from other cells in PBMC preparation.
scrapping is the best method used to subculture macrophages. scrappers are available with many manufacturers. gently scarpe the cells inside the culture flask in one direction. you can use a black paper as background to visualize the disadherence while scrapping. you should be very careful and scrapping should be done very slowly so that cells are not hurt or activated
all the best