Can anyone help by providing a protocol for splenic macrophage culture (preferably by plate adherence method)?
Does plate adherence for 2-3h or overnight work to get macrophages from mouse spleen?
Instead of using splenic macrophages (isolation by adherence), I used spleen-derived macrophages from C57BL/6 mice (cultured similarly as bone marrow-derived macrophages) following this protocol (instead of M-CSF, I also tried using L929 conditioned medium) and it worked.
J Immunol Methods. 2008 Sep 30;338(1-2):47-57.
An efficient culture method for generating large quantities of mature mouse splenic macrophages.
Alatery A, Basta S.
(PMID: 18675819)
As explained in the paper, spleen-derived macrophages do show some differences from bone marrow-derived macrophages. I hope this will help.
Another similar publication is:
Characterization of murine macrophages from bone marrow, spleen and peritoneum.
Wang C et al (BMC Immunol. 2013)
PMID: 23384230
Yes, adherence would work, though you are not going to get a pure population of macrophages, other adherent cells will be there. But if that's not an issue, the procedure is simple and effective. Just make sure you plate enough cells, since the majority will be non-adherent lymphocytes.
thanks for your responses... it is great help.. gonna start cultures on Mon..
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