When I stained mouse PBMCs, I saw there was a population of Ly6c+CD11b- cells (>10%) and these cells express CD86. From the FSC and SSC, I can tell they are not lymphocytes. Does anybody have an idea what kind of cell type these cells are? Enclosed is a figure from a JEM (2012) paper. They could also see this Ly6c+CD11b- population.
It could be a dendritic cell type that is Ly6c+CD11b-. This poster from R&D has a few cell types that could be what you are trying to identify. The mouse dermal DC population seems to fit the CD86+CD11b-Ly6c+ criteria, since DCs express CD86. But it would be hard to make a positive identification without a few more markers to be sure.
http://www.rndsystems.com/poster_detail_objectname_dendritic_cell_subsets.aspx
Another possibility is that they could be osteoprogenitor cells, as they are collected within the PBMC population and are CD11b-. Osteoblast progrnitor cells from the PBMC population can be CD86+, though I am not sure about Ly6C. But I would think that this population of cells in the PBMC is much lower than 10%.
Are these mice treated with anything special or do they have any infections?
These cells are CD11c negative. So they cannot be DCs. And they are from wild type mice without any treatment.
Hi Ken,
I am happy to look at your data and help figure out what it is, if you are interested. We have quite a bit of experience looking at myeloid populations in the mouse. We could do a shared dropbox folder. Email me at [email protected].
A quick run-down on surface markers on mouse leukocytes:
CD11b stains myeloid lineage cells: monocytes, neutrophils, eosinophils etc.
Ly6C is expressed on a whole lot of cell types. On CD11b+ cells, they are expressed on Ly6C(hi) monocytes, neutrophils, and eosinophils (to start with).
Here are my recommendations for gating these guys, so we can find out what it is:
1. Gate CD11b+ cells
2. Gate neutrophils (medium side scatter) and eosinophils (high side scatter), and monocytes (low side scatter).
Note: sometimes it's hard to separate neutrophils from monocytes on side-scatter alone. If you can't do this, then we can use the comparative levels of Ly6C to separate them.
3. The monocytes should separate into two populations -- Ly6C(hi) and Ly6C(lo). If the neutrophils have been included here, they are Ly6C(intermediate) and have slightly higher CD11b.
4. Eosinophils will have higher side scatter than neutrophils, and will be CD11b+Ly6C(int).
My recommendations for future staining:
- Include Ly6G as a specific marker for neutrophils.
- Eosinophils can then be gated using SSC vs Ly6G, as the are Ly6G- but higher side scatter.
could be a monocyte derived DC population at an intermediate stage of development ..
pDCs express low levels of CD11c (but they should be smaller still)
http://jem.rupress.org/content/suppl/2008/06/30/jem.20071365.DC1/2.html
you could include the following markers to get a better idea : MHCII, B220, PDCA-1; and maybe include a live dead marker as well.
to be extra sure include markers for T cells , NK cells in a common dump channel .
these cells should be monocytes (CD11B+Ly6Chi) and neutrophils (CD11B+Ly6C med). one can differentiate these two by Ly6G staining. Neutrophils are Ly6g+ and monocytes are -
I am using Ly6G for In vivo Biotinylation of mouse Neutrophil turnover kinetics . will it perfectly work or not.
They seem to be common monocyte-macrophage progenitors (CD11b neg Ly6C+). I observed this population of cells as well in the spleen of mice.
Here a link to a book which explains that.
https://books.google.fr/books?id=Q-e9CwAAQBAJ&pg=PA29&lpg=PA29&dq=CD11bneg+Ly6C%2B&source=bl&ots=ua_IUjW2Ea&sig=ACfU3U3sCUvobKuftMPSCUpwtah0HZPuuA&hl=fr&sa=X&ved=2ahUKEwju8JjYrebiAhUP3xoKHcunDTQQ6AEwAXoECAkQAQ#v=onepage&q&f=false
Best,
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