Given the problem of priming or pre-activation of isolated neutrophils:
Has anyone a suggestion (ideally first hand experience) for a good protocol injecting neutrophils into mice in order to perform short term experiments (max. 12h)
We would really prefer going for neutrophil depletion or competition experiments (i.e. without prior depletion) and not adoptive bone marrow transfer since we critically need to distinguish neutrophil from macrophage function in our read-out.
You should write clearly what to do? Which neutrophil will be used and what is the purification system. The root of injection should be mention. Whether is intra peritoneal Intra venously etc..
What is you experimental setup? We struggled over 1 year with the experimental setup. Depends how to do it. There is a protocol for neutrophil transfer in Köffel et al. 2014 (Blood, attached). However, few suggestion: FACS sorting of Neutrophils before transfer is not recommded as most of them die off after transfer. For injection i.v. you will have to inject "tons" of cells as most of them will get lost somewhere in the mouse (question if this is in any kind "physiological"))..............You could contact me directly via [email protected] to dissuss in detail.
Article Monocytic cell differentiation from band-stage neutrophils u...
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