Hi,
I'm looking for characterizing monocytes in human blood from dialysis patients and i used CD14 CD16 as a marker for the monocytes subunits out of the leukocyte gate. however, the percentage of CD14 is too low. is there a better marker or a better strategy for detecting monocytes.
I would love to hear some suggestions
IMMUNOLOGY
FLOW CYTOMETRY
MONOCYTES
HI there,
well, too low by what measure? Cell counts in individuals can vary quite widely, and may also vary according to time of day, when they ate, do they have a sub clinical infection and a whole bunch of things (such as when did you measure it relative to their last dialysis).
What sort of numbers did you get from other dialysis patients before and after dialysis, and from healthy people?
Good Luck!
G
Hi Miran
Grant is right. Monocytes are 2-10% of white blood cell count.
CD14high is the best marker for monocyte. However it has been reported CD14low monocytes and also downregulation of CD14, both due to some illness.
As for CD16, not all monocytes express CD16. There are CD16pos and CD16neg monocytes. So if you are gating on CD14posCD16pos your cell percentage might be lower than expected.
CD33 is also a possible marker but you would need to gate out neutrophils (CD66b) and DCs (CD11c).
http://intimm.oxfordjournals.org/content/13/12/1571.full
http://www.ncbi.nlm.nih.gov/pubmed/22837127
http://www.ncbi.nlm.nih.gov/pubmed/17523943
Hi Miran
Darrell et.al 2009 reported ( identified markers that discriminate between human peripheral blood monocytes, tissue macrophages, fibrocytes, and fibroblasts. Amongst these four cell types, only peripheral blood monocytes express the combination of CD45RO, CD93, and S100A8/A9; only macrophages express the combination of CD45RO, 25F9, S100A8/A9, and PM-2K; only fibrocytes express the combination of CD45RO, 25F9, and S100A8/A9, but not PM-2K; and only fibroblasts express the combination of CD90, cellular fibronectin, hyaluronan, and TE-7. These markers are effective both in vitro and in sections from human lung. We found that markers such as CD34, CD68, and collagen do not effectively discriminate between the four cell types. In addition, IL-4, IL-12, IL-13, IFN-γ, and SAP differentially regulate the expression of CD32, CD163, CD172a, and CD206 on both macrophages and fibrocytes. Finally, CD49c (α3 integrin) expression identifies a subset of fibrocytes, and this subset increases with time in culture.
thank you guys for your answers.
but, Is cd 64 out of cd 45 a good marker for monocytes?
CD45 is expressed by all white blood cells, I think CD64 is expressed on all monocytes (intensity can vary it's a marker of activation)
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