Induction, Isolation, and Characterization of Human Neutrophil Extracellular Traps (NETs) to Be Used in Functional...

Polymorphonuclear neutrophils (PMNs) represent ~60% of circulating leukocytes in humans. Once activated, PMNs can release neutrophil extracellular traps (NETs), molecular complexes composed of DNA and proteins from granules. Although NETs play a protective role in fighting microbes, they may become harmful under certain circumstances, especially in some inflammatory autoimmune diseases.Because NETs can be either beneficial or deleterious depending on the microenvironment, a deeper characterization of NETs and NET-inducing stimuli as well as a better understanding of the different properties and activities of NETs is required, in both physiological and pathological contexts.In the present chapter, we describe tools not only to induce and isolate NETs, but also required methods to characterize NETs via biochemical approaches and imaging, in order to prove that obtained structures conform to the definition of NETs exhibiting typical extracellular fibers. NETs produced with our protocol are suitable for detailed assessment of their biological properties in functional studies.