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Isolation of Human Neutrophils from Whole Blood and Buffy Coats
Published on: September 17, 2021
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Isolation and Functional Analysis of Human Neutrophils
Douglas B Kuhns1, Debra A Long Priel1, Jessica Chu2
1Neutrophil Monitoring Lab, Leidos Biomedical Research Inc., Frederick National Laboratory for Cancer Research, Frederick, Maryland.
Current Protocols in Immunology
|November 4, 2015
Summary
This study details isolating human polymorphonuclear neutrophils (PMN) from blood using density gradient methods. It also outlines key assays for evaluating neutrophil functions like respiratory burst and phagocytosis.
Area of Science:
- Immunology
- Cell Biology
- Hematology
Background:
- Human polymorphonuclear neutrophils (PMN) are critical immune cells.
- Efficient isolation and functional assessment of PMN are essential for immunological research.
Purpose of the Study:
- To describe a standardized method for isolating human PMN from peripheral blood.
- To outline functional assays for evaluating key neutrophil activities.
Main Methods:
- Isolation of PMN using dextran sedimentation followed by density gradient centrifugation (Percoll or Ficoll-Paque).
- Functional assays including measurement of respiratory burst activation, phagocytosis assays, and microbial killing assays.
Main Results:
- Successful isolation of viable human PMN with high purity.
- Demonstration of functional integrity of isolated PMN through various assays.
Conclusions:
- The described methods provide a reliable approach for obtaining human PMN for research.
- These techniques facilitate the study of neutrophil function in various physiological and pathological conditions.
