Question

To separate macrophages from a mouse

Answer 1

You should not need to use Ficoll for mouse blood, instead we just ensure RBC lysis is complete - we use Biolegend RBC lysis buffer, but any should do. Following this block FC receptors with anti-CD16/32. Then stain with anti CD115, APC works well for me. If you have the option to flow sort, then you can further separate these cells on Ly6C surface expression (I would recommend a lineage gate to remove T cells, B-cells and granulocytes). If not, then you could use an anti-APC bead selection kit to pull out your monocytes

Answer 2

Previous studies have shown the presence of human macrophages and dendritic cells in the humanized mice. ... Although human RBCs became detectable in humanized mice after macrophage depletion, the levels of human RBC chimerism were still significantly lower than human PBMC chimerism.

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