I have generated DC from mice bone marrow. I have followed very known protocol, incubating with GM-CSF for 8 days and LPS activation overnight or 48 hours.
After LPS activation, I stained my cells with CD11b, MHC II and CD 11C and then FACS analysing to be sure about DC population.
Results: I have around 30% CD 11b low, and 70% cd 11b high.
From gated CD 11b hi cells --> 70 % MHC II hi CD11 C + (activated DC) while 30% are MHC II hi CD11 -/low.
So I am wondering what are those CD11C low and MHC hi cells? Are they inflammatory monocytes (M1)? Or they are the precursor of CD11 hi cells?
The dendritic cells are not all CD11c +, far from it.
Most DCs which are CD205 + and / or 33D1 + are also CD11c-.
It is the same for Langerin + and + DCLamp.
It is therefore quite possible to have Class II + and CD11c - or low DCs.
On the other hand, the use of LPS in the presence of GM-CSF, reverses the ability to have real DCs.
It would be more judicious to use cytokines such as TNFα, IL15, FLT3L, IL4 or IL10, CD154 activator ... rather than LPS which is an inhibitor of DC development.
Those MHCII low DCs are probably DCs that are less activated than their MHCII high counterparts. To be sure you could check the expression levels of other markers such as CD80 or CD86. I think that this article could also help you: https://www.ncbi.nlm.nih.gov/pubmed/26084029
Jean and Jorge @ Thanks a lot
Binnur @ Are what types of macrophages they might be? What about inflammatory monocytes?
Do you know any paper or review paper that investigated it in more detail? Thanks a lot
Dendritic cells cells. See paper:
Article Phenotypic Characterization of Five Dendritic Cell Subsets i...
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