Zika Virus Infectious Cell Culture System and the <em>In Vitro</em> Prophylactic Effect of Interferons

Deisy Contreras; Vaithilingaraja Arumugaswami

doi:10.3791/54767

Virus Zika sistema de cultura celular Infecciosas e do In Vitro Efeito profilático de in...

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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 de cultura celular Infecciosas e do In Vitro Efeito profilático de interferões

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09:11 min

August 23, 2016

DOI: 10.3791/54767-v

Deisy Contreras¹, Vaithilingaraja Arumugaswami^1,2,3

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.

O vírus Zika (ZIKV), um patógeno emergente, está ligado a anormalidades no desenvolvimento fetal e microcefalia. O estabelecimento de um sistema eficaz de cultura de células infecciosas é crucial para estudos de replicação do ZIKV, bem como para o desenvolvimento de vacinas e medicamentos. Neste estudo, vários ensaios virológicos relativos ao ZIKV são ilustrados e discutidos.

O objetivo geral deste estudo é estabelecer um sistema de cultura infecciosa in vitro do vírus Zika baseado em células de mamíferos para estudar a replicação do vírus Zika, bem como a utilização da plataforma in vitro para o desenvolvimento de medicamentos. Este protocolo detalhado passo a passo do sistema de cultura de células infecciosas do vírus Zika pode ajudar a abordar o mecanismo da patogênese da doença mediada pelo vírus Zika e a evolução da virulência viral. A principal vantagem dessa técnica é que ela pode permitir a produção em larga escala do vírus Zika para o desenvolvimento de medicamentos e vacinas.

Comece colhendo células Vero com 80% de densidade celular em um frasco de cultura T75, aspirando o meio do frasco e enxaguando as células com dois mililitros de solução salina tamponada com fosfato. Adicione dois mililitros de 0,25% de tripsina e incube o frasco a 37 graus Celsius por cinco minutos. Após a incubação, adicione oito mililitros de meio de crescimento contendo soro para inativar a tripsina.

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