Formación de trampas extracelulares de neutrófilos humanos in vitro: descripción de un método

Lizet Jazmín Pérez Zapata; Ana María Trejos Ramírez; Andres Augusto Arias Sierra; Juan Álvaro López Quintero

doi:10.17533/udea.hm.v10n1a01

Authors

Lizet Jazmín Pérez Zapata University of Antioquia
Ana María Trejos Ramírez University of Antioquia
Andres Augusto Arias Sierra University of Antioquia
Juan Álvaro López Quintero University of Antioquia

DOI:

https://doi.org/10.17533/udea.hm.v10n1a01

Keywords:

standardization, neutrophils, NETs, Phorbol 12-myristate 13-acetate, extracellular neutrophil traps, netosis

Abstract

Introduction: Neutrophil Extracellular Traps (NET) are net-shaped structures, composed of DNA and granule proteins, which are released by neutrophil into the extracellular space in response to different stimuli. NETosis is described as a microbicide mechanism, which destroys microorganisms or inhibits their growing.

Objective: To describe a process to induce, visualize and quantify the formation of NET in human neutrophils.

Methodology: peripheral blood neutrophils were isolated from healthy individuals. NET production was induced using different concentrations of PMA at specific times. DNA release into the extracellular space was quantified by spectrofluorimetry and NET morphology was evaluated by fluorescence microscopy. Statistical analyzes were performed using SPSS and GraphPad Prism programs.

Results and conclusions: this study describes a method used to induce NET in human neutrophils, which is comparable with other studies reported in the scientific literature. These results can be applied in essays that seek to evaluate NET formation in vitro and expect it will be helpful to researchers who wish to study this cellular process.

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Author Biographies

Lizet Jazmín Pérez Zapata, University of Antioquia

Primary Immunodeficiencies Group, Faculty of Medicine, University of Antioquia.

Ana María Trejos Ramírez, University of Antioquia

Primary Immunodeficiencies Group, Faculty of Medicine, University of Antioquia.

Andres Augusto Arias Sierra, University of Antioquia

Primary Immunodeficiencies Group, Faculty of Medicine, University of Antioquia.

Juan Álvaro López Quintero, University of Antioquia

Primary Immunodeficiencies Group, Faculty of Medicine, University of Antioquia.

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