I was wondering what is the best way to isolate neutrophils from the bone marrow without activation.
what is the approximate number of cells will I get from one mouse 8-12 weeks?
there is a couple of methods to do isolation.
with percoll gradient you can get around 6million cells per mouse
I am looking into immunomeganetic beads from Miltenyi and Stemm Cell technologies, but I don't know the numbers of
Please find the protocol in our bellow paper mentioning BMDN isolation from mouse femoral bones:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4345018/
Are you looking to get purified (or enriched) neutrophils out from the marrow, or do you just want to identify the neutrophils?
From mouse femoral bones you can get 10 million cells per mouse, when using MACS GR1+ positive selection at around 97% purity, alternatively you can use Ly6G+ selection. Depending on your experimental needs you could also run a negative selection for "untouched" neutrophils but for most neutrophil functional assays Gr1+ selection is fine. Contact me if you need more details for isolating neutrophils.
Make sure to not activate the neutrophils during the selection process. You can find nice protocols in pubmed by searching for Laskay-T
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