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Updated: Jul 16, 2025
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Isolation, Purification and Labeling of Mouse Bone Marrow Neutrophils for Functional Studies and Adoptive Transfer Experiments
Published on: July 10, 2013
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Isolation of Mouse Neutrophils
Andrew L Wishart1, Muthulekha Swamydas1, Michail S Lionakis1
1Fungal Pathogenesis Section, Laboratory of Clinical Immunology & Microbiology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA.
Current Protocols
|September 14, 2023
Summary
This study provides standardized protocols for isolating neutrophils, crucial immune cells, from mouse bone marrow and tissues. These methods ensure high purity and viability for studying neutrophil function in infection and immunity.
Area of Science:
- Immunology
- Cell Biology
- Hematology
Background:
- Neutrophils are vital for innate immunity against bacterial and fungal infections.
- Defects in neutrophil number or function increase susceptibility to severe infections.
- Understanding neutrophil function is key to developing treatments for infectious diseases.
Purpose of the Study:
- To describe reproducible methods for isolating high-purity, viable neutrophils from mouse bone marrow and tissues.
- To provide standardized protocols applicable in diverse laboratory settings.
- To enable detailed ex vivo analysis of neutrophil functions and adoptive transfer experiments.
Main Methods:
- Density-gradient centrifugation for neutrophil isolation from bone marrow.
- Positive and negative immunomagnetic selection for bone marrow neutrophil purification.
- Enzymatic tissue digestion combined with immunomagnetic selection or FACS for tissue neutrophil isolation.
Main Results:
- Established protocols yield large numbers of highly enriched and viable neutrophils.
- Methods are effective for isolating neutrophils from mouse bone marrow, spleen, liver, and kidney.
- Isolated neutrophils are suitable for functional assays including phagocytosis, oxidative burst, and cytokine production.
Conclusions:
- These validated protocols facilitate robust research into neutrophil biology and function.
- Standardized isolation techniques are essential for reproducible studies in immunology and infectious disease.
- The methods support investigations into neutrophil-mediated immunity and the development of novel therapeutic strategies.
