In Vitro Identification and Isolation of Human Neutrophil Extracellular Traps

Methods Mol Biol. 2021:2255:97-117. doi: 10.1007/978-1-0716-1162-3_10.

Abstract

Neutrophils release web like-structures known as neutrophil extracellular traps (NETs) that ensnare and kill microorganisms. These networks are constituted of a DNA scaffold with associated antimicrobial proteins, which are released to the extracellular space as an effective mechanism to fight against invading microorganisms. In parallel with this beneficial role to avoid microbial dissemination and wall off infections, accumulating evidence supports that under certain circumstances, NETs can exert deleterious effects in inflammatory, autoimmune, and thrombotic pathologies. Research on NET properties and their role in pathophysiological processes is a rapidly evolving and expanding field. Here, we describe a combination of methods to achieve a successful in vitro NET visualization, semiquantification, and isolation.

Keywords: Cell death; Elastase; Myeloperoxidase; NETosis; Neutrophil extracellular traps.

MeSH terms

  • Cell Separation / methods*
  • DNA / analysis*
  • Extracellular Traps / metabolism*
  • Humans
  • Image Processing, Computer-Assisted / methods*
  • In Vitro Techniques
  • Microscopy, Fluorescence / methods*
  • Pancreatic Elastase / analysis*
  • Peroxidase / metabolism*

Substances

  • DNA
  • Peroxidase
  • Pancreatic Elastase