FACS Isolation of Macrophages from Mouse

[proatpcr]

Hi im new to FACS and macrophages. I need to figure out how to isolate these guys from mouse liver and spleen especially from Kupffer cells in order to do some apoptotic and phagocytic assays using FACS.

Can i use Miltenyi CD11b beads to do the isolation, but does this not affect their function?

thanks for your help

[IoDiNE]

These methods will work fine for isolating monocytes from the spleen and function usually is fine

I have also used the plating method to isolate away contaminating monocytes.

You can leave the suspension for 2-3 hours but others leave it overnite. In order to remove cells from the plastic you can use straight pipetting force for RAW cells and if they are very sticky also 0.53 mM EDTA in PBS works fine to remove them.

I have been using monocytic suspension cell culture in order to assay cytokines and do not differentiate them to allow easier analysis by flow cytometry.

cheers