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JoVE Journal
Immunology and Infection
Virus Zika Sistema infecciosa cultivo celular y la In Vitro Profiláctica Efecto de los i...
Virus Zika Sistema infecciosa cultivo celular y la In Vitro Profiláctica Efecto de los i...
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Zika Virus Infectious Cell Culture System and the In Vitro Prophylactic Effect of Interferons

Virus Zika Sistema infecciosa cultivo celular y la In Vitro Profiláctica Efecto de los interferones

Full Text
21,558 Views
09:11 min
August 23, 2016

DOI: 10.3791/54767-v

Deisy Contreras1, Vaithilingaraja Arumugaswami1,2,3

1Board of Governors Regenerative Medicine Institute,Cedars-Sinai Medical Center, 2Department of Surgery,Cedars-Sinai Medical Center, 3Department of Surgery, David Geffen School of Medicine,University of California, Los Angeles

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Overview

This study establishes a mammalian cell-based in vitro culture system for Zika virus (ZIKV) to investigate its replication and facilitate drug development. The detailed protocol aims to enhance understanding of ZIKV pathogenesis and support large-scale virus production for therapeutic purposes.

Key Study Components

Area of Science

  • Virology
  • Infectious disease research
  • Cell culture techniques

Background

  • Zika virus is associated with severe fetal developmental issues.
  • Effective culture systems are essential for studying ZIKV.
  • Understanding ZIKV replication aids in vaccine and drug development.
  • Current methods lack efficiency for large-scale production.

Purpose of Study

  • To establish an infectious cell culture system for ZIKV.
  • To facilitate studies on ZIKV replication mechanisms.
  • To support drug and vaccine development efforts.

Methods Used

  • Harvesting Vero cells at 80% density in T75 flasks.
  • Rinsing cells with phosphate-buffered saline.
  • Incubating with trypsin to detach cells.
  • Adding growth medium to inactivate trypsin.

Main Results

  • Successful establishment of an in vitro ZIKV culture system.
  • Detailed protocol for cell culture and virus production.
  • Potential for large-scale ZIKV production demonstrated.
  • Insights into ZIKV pathogenesis and virulence evolution.

Conclusions

  • The established system is a valuable tool for ZIKV research.
  • It enhances understanding of ZIKV disease mechanisms.
  • Supports future drug and vaccine development initiatives.

Frequently Asked Questions

What is the significance of Zika virus research?
Zika virus research is crucial due to its association with severe fetal abnormalities and the need for effective vaccines and treatments.
How does the in vitro culture system aid in ZIKV studies?
The in vitro culture system allows researchers to study ZIKV replication and pathogenesis in a controlled environment, facilitating drug and vaccine development.
What cell line is used in the study?
Vero cells are used for establishing the ZIKV infectious culture system.
What are the main advantages of this technique?
The technique allows for large-scale production of ZIKV, which is essential for research and therapeutic development.
What protocols are included in the study?
The study includes detailed protocols for harvesting cells, detaching them, and preparing the culture system for ZIKV infection.

El virus del Zika (ZIKV), un patógeno emergente, está relacionado con anomalías del desarrollo fetal y microcefalia. El establecimiento de un sistema eficaz de cultivo de células infecciosas es crucial para los estudios de replicación del ZIKV, así como para el desarrollo de vacunas y fármacos. En este estudio, se ilustran y discuten varios ensayos virológicos relacionados con el ZIKV.

El objetivo general de este estudio es establecer un sistema de cultivo infeccioso del virus del Zika basado en células de mamíferos para estudiar la replicación del virus del Zika, así como la utilización de la plataforma in vitro para el desarrollo de fármacos. Este protocolo detallado paso a paso del sistema de cultivo de células infecciosas del virus del Zika puede ayudar a abordar el mecanismo de la patogénesis de la enfermedad mediada por el virus del Zika y la evolución de la virulencia viral. La principal ventaja de esta técnica es que puede permitir la producción a gran escala del virus del Zika para el desarrollo de fármacos y vacunas.

Comience cosechando células Vero al 80% de densidad celular en un matraz de cultivo T75 aspirando el medio del matraz y enjuagando las células con dos mililitros de solución salina tamponada con fosfato. Añadir dos mililitros de tripsina al 0,25% e incubar el matraz a 37 grados centígrados durante cinco minutos. Después de la incubación, agregue ocho mililitros de medio de crecimiento que contenga suero para inactivar la tripsina.

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