Hello everyone. I have collected intestinal biopsies from stem cell transplanted patients in RNAlater and wish to quantify neutrophils only via qPCR. As we know that the mucosa of gut is filled with several immune/epithelial cells, I want to target just one cell population. Is there any singular magic marker for neutrophils? Please let me know. Thank you in advance.
Hi,
I would recommend a meta-analysis of leukocytes in atherosclerotis mouse aorta by single cell sequencing, which allows for the identification of different immune cells as attached. In this paper, they recommend to use C7d, S100a9 and S100a8 to identify neutrophils instead of canonical markers. Indeed, you could look at the top gene signature of each clusters in Figure 4, you would find 10 genes specifically expressed in neutrophils and rarely expressed in other immune cell types.
Hope it helps!
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Thank you Yi Yan for your input...... I will of course think about it. This paper is in the context of mouse. I have over 500 human patient sample waiting for qPCR. Therefore, I am reluctant to use plural genes. With those many genes, I may bring my bed to the lab and eat and sleep here :D. If you were to recommend me one gene for human neutrophil, what would you recommend me?
Hi Sakhila,
according to this paper, I would suggest either S100a8 or S100a9. Actually, you can order both primers (the price is not high) and run 3 human samples for a test to check whether the primer works or not instead of running all the sample at one time. Then you can finally decide the primer to go for 500 human patient.
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