Hello,
has anybody a good protocol for the isolation of macrophages from the large intestine?
I have one but the cells I get are not very adherent... The basics of this protocol are the isolation of the lamina propria cells, F4/80 Isolation via MACS and these cells I used for FACS or took them in culture.
Questions I have:
- Is maybe a Percoll a better option for the purity instead of the F4/80 Isolation? Because the macrophages get adherent after 2h, so I could wash all the others cells away, but also the fibroblasts get adherent which would be a problem (I dont´t want a co-culture)....
- should I also use the IEL fraction and not only the LP fraction for macrophage isolation?
I would be very grateful if anybody has some experience and could give some advice :)
Hi, so the IEL cells would be T cells. I think the best way is to do the Lamina Propria and then you can put them in the magnetic columns with CD45 beads.
Then, you stain for the macrophages. Should work well.
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