I am currently using the Mouse Neutrophil Negative Selection Kit (HY-K0350) to isolate neutrophils from C57BL/6 mice. I would like to inquire about the expected yield of neutrophils from both mouse bone marrow and peripheral blood. How many neutrophils can typically be obtained from each source? Has anybody tried this before? Any insights or experiences regarding this would be greatly appreciated!
Hi Marine Ma
The yield of cells can vary greatly depending on mouse strain, age, and other factors, so it’s difficult to give an exact number. Since neutrophils account for only a limited proportion of peripheral blood (about 10–25%), and blood volume also differs significantly with age (adult versus juvenile mice may differ by several fold), the final yield is also heavily influenced by sample preprocessing. Therefore, neutrophil yield from peripheral blood is unlikely to reach the million-cell level. Given this variability, it is generally recommended to use bone marrow samples as a more reliable source for experiments.
The answer to this question is provided by MedChemExpress Technical Support.
Hello Marine Ma,
From an adult mouse's bone marrow (two femurs and two tibias), you can expect to isolate 6-12 million viable neutrophils using method like density gradient centrifugation.
You may want to refer to the article attached below for more information.
https://pmc.ncbi.nlm.nih.gov/articles/PMC10503263/
Highlight:
The typical yield of cells from the bone marrow (i.e., 2 femurs and 2 tibias) of an uninfected 8-12 week-old C57BL/6 mouse is between 60 and 80 million. The typical yield of neutrophils from the bone marrow following positive immunomagnetic selection is ~7-12 million neutrophils with >97% purity and >97% viability as determined by flow cytometry. The typical yield of neutrophils from the bone marrow following negative immunomagnetic separation is ~4-6 million neutrophils with >91% purity and >98% viability. For Histopaque-based gradient centrifugation method for bone marrow neutrophil isolation, the typical yield is ~6-12 million neutrophils with >91% purity and >98% viability.
However, the yield from mouse peripheral blood is considerably lower, as neutrophils only make up about 10-20% of total white blood cells.
You may want to refer to the articles attached below for more information.
https://pmc.ncbi.nlm.nih.gov/articles/PMC5518602/
https://www.researchgate.net/publication/384814278_Proteomic_characterization_of_1000_human_and_murine_neutrophils_freshly_isolated_from_blood_and_sites_of_sterile_inflammation
Best,