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Morphological and Compositional Analysis of Neutrophil Extracellular Traps Induced by Microbial and Chemical Stimuli
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1Clinical Immunology Research Department of Biochemical Sciences Faculty,Universidad Autónoma “Benito Juárez” de Oaxaca, 2Department of Molecular Biomedicine,Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, 3CONACYT- Medicine and Surgery Faculty,Universidad Autónoma “Benito Juárez” de Oaxaca, 4Molecular Immunology Research Department of Medicine and Surgery Faculty,Universidad Autónoma “Benito Juárez” de Oaxaca
Capítulos
- 00:05Introduction
- 01:35Peripheral Blood Collection and Polymorphonuclear Neutrophil Purification
- 04:11Neutrophil Morphology and Viability
- 07:28CFSE Staining of Microorganisms and NET Induction
- 11:10Results: Analysis of NET Induced in Human Blood Derived Neutrophils
- 13:19Conclusion
Presented here is a protocol for the induction and analysis of in vitro neutrophil extracellular traps (NETs). Quantification of DNA, cathelicidin (LL37), and enzyme activity yielded data that show the variability in the composition and morphology of NETs induced by microbial and chemical stimuli under similar controlled conditions.
