Formación de trampas extracelulares de neutrófilos humanos in vitro: descripción de un método
DOI:
https://doi.org/10.17533/udea.hm.v10n1a01Palabras clave:
estandarización, neutrófilos, NETs, netosis, Phorbol 12-myristate 13-acetate, trampas extracelulares de neutrófilosResumen
Introducción: las trampas extracelulares de neutrófilo (NETs) son estructuras en forma de red compuestas de ADN y proteínas granulares que el neutrófilo libera al espacio extracelular en respuesta a diferentes estímulos. La NETosis es un mecanismo de muerte celular, que además cumple una función microbicida destruyendo o inhibiendo el crecimiento de microorganismos.
Objetivo: describir el proceso para inducir, visualizar y cuantificar la formación de NETs en neutrófilos humanos.
Métodos: se aislaron neutrófilos de sangre periférica de individuos sanos y se indujo la producción de NETs utilizando diferentes concentraciones de PMA a determinados tiempos. La liberación de ADN se cuantificó por espectrofluorimetría y se evaluó la morfología de las NETs utilizando microscopía de fluorescencia. Los análisis estadísticos se realizaron con los programas SPSS 17.02 y GraphPad Prism 6.
Resultados y conclusiones: en el presente trabajo se describe un método que sirve para inducir NETs utilizando neutrófilos humanos, el cual es comparable con otros estudios descritos en la literatura. Los resultados y técnicas descritas aquí se pueden aplicar en ensayos que busquen evaluar el proceso de formación de NETs in vitro y tiene como intensión brindar herramientas metodológicas a los investigadores que deseen estudiar este proceso celular.
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