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Related Experiment Videos

Effects of different density gradient separation techniques on neutrophil function

T J Venaille1, N L Misso, M J Phillips

  • 1Department of Medicine, University of Western Australia, Nedlands.

Scandinavian Journal of Clinical and Laboratory Investigation
|August 1, 1994
PubMed
Summary
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Percoll gradients may be the preferred method for isolating polymorphonuclear leukocytes (PMNs) due to minimal cell activation during separation. This technique offers a quick and effective way to obtain pure, functional PMNs for research.

Area of Science:

  • Immunology
  • Cell Biology

Background:

  • Investigating inflammatory disease pathogenesis requires assessing polymorphonuclear leukocyte (PMN) activity.
  • Various methods exist for separating PMNs from whole blood for research purposes.

Purpose of the Study:

  • To compare the effects of three PMN separation techniques: Percoll gradient, Ficoll-Hypaque/Dextran sedimentation (FH/DS), and Mono-Poly Resolving Medium (M-PRM).
  • To evaluate the impact of each method on PMN activation and function.

Main Methods:

  • Comparison of Percoll gradient, FH/DS, and M-PRM for PMN separation.
  • Assessment of cell yield, purity, pseudopodia formation, and resting superoxide release.
  • Evaluation of functional responses including chemotaxis and stimulated superoxide production.

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Main Results:

  • All methods yielded comparable cell purity and quantity.
  • Percoll separation showed lower spontaneous pseudopodia formation and superoxide release compared to FH/DS and M-PRM.
  • Functional capacity, assessed by chemotaxis and stimulated superoxide production, was similar across all methods.

Conclusions:

  • While all three methods yield functionally active PMNs, Percoll gradients may be advantageous.
  • Percoll gradients offer a rapid separation method with minimal pre-activation of PMNs.
  • This makes Percoll a potentially preferable choice when preserving PMN resting state is critical.