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Updated: Jul 5, 2026
Isolation, Purification and Labeling of Mouse Bone Marrow Neutrophils for Functional Studies and Adoptive Transfer Experiments
Published on: July 10, 2013
Isolation of mouse neutrophils
1Harvard Medical School, Boston, Massachusetts.
Abstract:
Neutrophils are widely used in research to elucidate the mechanisms of inflammation, including such processes as cell adhesion, chemotaxis, superoxide release, production of reactive nitrogen intermediates, and granule exocytosis. This unit presents a protocol for the induction of an enriched exudate of polymorphonuclear leukocytes (PMN) in the peritoneal cavity of mice. Neutrophils are then isolated using continuous gradient centrifugation. This is followed by a protocol for isolating neutrophils from peripheral blood. An alternate method for the purification of PMN from peritoneal exudate cells or peripheral blood by Histopaque density gradient centrifugation is also provided. The cell purification protocol can be modified to obtain nonelicited neutrophils from peritoneal fluid.
Insights
This study details methods for isolating neutrophils, a key cell type in inflammation research. Protocols are provided for obtaining enriched neutrophils from mouse peritoneal cavities and peripheral blood for further study.
Area of Science:
- Immunology
- Cell Biology
- Biomedical Research
Background:
- Neutrophils are critical immune cells involved in inflammatory processes.
- Understanding neutrophil function requires reliable methods for their isolation and purification.
- Existing methods may require optimization for specific research applications.
Purpose of the Study:
- To present a comprehensive protocol for isolating neutrophils from mouse models.
- To offer methods for obtaining enriched populations of polymorphonuclear leukocytes (PMN).
- To provide alternative purification techniques for diverse research needs.
Main Methods:
- Induction of PMN exudate in the mouse peritoneal cavity.
- Isolation of neutrophils via continuous gradient centrifugation.
- Purification of neutrophils from peripheral blood and peritoneal exudates using Histopaque density gradients.
Main Results:
- Successful isolation of enriched neutrophil populations from mouse peritoneal cavities.
- Established protocols for neutrophil isolation from peripheral blood.
- Demonstrated adaptability of purification methods for non-elicited neutrophils.
Conclusions:
- The presented protocols provide robust methods for obtaining purified neutrophils for research.
- These techniques facilitate the study of neutrophil functions in inflammation.
- The protocols are adaptable for various experimental designs requiring neutrophil isolation.
