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> We have an investigator who wants to look at mouse dendritic cells
> in a chimeric model. Since there are no commercial dendritic markers
> available, we are using a combination of I-A , size, and exclusion of B
> cells and macrophages to find these cells. Any suggestions? Has anyone
> tried to identify mouse dendritic cells by flow.
The Lymphocyte Differentiation Unit here at WEHI has worked
extensively with dendritic cells (mouse and human) over several years.
In order to study them in isolation, the method invariably used these
days is first to isolate them (from mouse or human spleen or thymus,
human blood or tonsil) using density followed by magnetic bead
separation (for mouse, negative separation using anti- CD3, CD4,
Thy-1, IL2-R, Gr-1, Mac-1, macrophage F4/80, B220, TER119, FcR II)
(See David Vremec et. al. J. Exp. Med. 176, 47-58 1992 and references
therein). These are then 75-95% pure. They can be FACS sorted to
purity using class II MHC *or* CD8 for which two receptors they are
very bright. They bear "dendritic cell" markers 33D1, NLDC145 and
MTS35 (the latter is also on some lymphoid cells), and also several
T-cell (and other) markers.
Hope this helps.
Regards,
\ / < Flow Systems Laboratory
Frank Battye \__/ <<<<< The Walter & Eliza Hall Institute
Robyn Muir ----------!!<<<<<<<< Voice: 61_3_9345 2540
Dora Constantinou /!!\ <<<<< Fax: 61_3_9347 0852
o !! \ < IN:: "facs_copy@wehi.edu.au"
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