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Biologia
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Isolation of Human Neutrophils from Whole Blood and Buffy Coats
JoVE Journal
Biologia
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JoVE Journal Biologia
Isolation of Human Neutrophils from Whole Blood and Buffy Coats

Isolation of Human Neutrophils from Whole Blood and Buffy Coats

DOI:
10.3791/62837-v

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08:12 min

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September 17, 2021

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, , ,
1Department of Pathology,Harvard Medical School, Department of Lab Medicine, Children’s Hospital Boston, and Dana-Farber/Harvard Cancer Center, 2Department of Microbiology, Faculty of Science,Mahidol University

Capítulos

  • 00:04Introduction
  • 00:45Gradient Layering
  • 02:40Erythrocyte Sedimentation
  • 03:45Residual RBC Lysis
  • 05:06Quality Determination of Isolated Neutrophil
  • 05:44Results: Neutrophil Yield and Quality Estimation
  • 07:34Conclusion

Summary

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This protocol details a method for the isolation of neutrophils from whole blood, buffy coats, or leukapheresis membranes, achieving good yield, high purity, and minimal cell activation. We utilize gradient purification, red blood cell (RBC) sedimentation, and RBC lysis to obtain a high-quality/purity neutrophil preparation.

Tags

Keywords: NeutrophilsDensity GradientSedimentationLysisBuffy CoatPBMCErythrocyteCell BiologyCell IsolationCell Preparation

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