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Updated: Jun 27, 2026
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Neutrophil Isolation Protocol
Published on: July 23, 2008
Neutrophil isolation protocol.
Hana Oh1, Brian Siano, Scott Diamond
1Institute for Medicine and Engineering, University of Pennsylvania, USA. hanaoh@seas.upenn.edu
Journal of Visualized Experiments : Jove
|December 11, 2008
Summary
This study presents a density gradient method for isolating human neutrophils (polymorphonuclear granulocytes) from blood. The technique yields highly pure and viable neutrophils for essential in vitro research.
Area of Science:
- Immunology
- Cell Biology
- Hematology
Background:
- Neutrophils (polymorphonuclear granulocytes) are critical immune cells involved in inflammation.
- Their functions, including locomotion and phagocytosis, require pure cell populations for accurate study.
- Short neutrophil lifespan necessitates rapid isolation for timely in vitro experiments.
Purpose of the Study:
- To establish a reliable and efficient method for isolating human neutrophils from whole blood.
- To provide researchers with a practical technique for obtaining high-purity neutrophil samples for functional assays.
Main Methods:
- Utilized a standard density gradient separation using a sodium metrizoate and Dextran 500 mixture.
- Involved layering whole blood over the gradient medium, followed by centrifugation.
- Included separation of the neutrophil layer and subsequent lysis of erythrocytes.
Main Results:
- Achieved high purity, with isolated samples containing >95% neutrophils.
- Maintained high cell viability (>95%) in the isolated neutrophil preparations.
- Demonstrated a fast and reproducible method suitable for short-lived neutrophils.
Conclusions:
- The described density gradient method effectively isolates pure, viable human neutrophils.
- This technique supports critical in vitro studies of neutrophil function in inflammation and immunity.
- The method is practical for researchers needing rapid access to neutrophils within hours of blood collection.
