Dados do Trabalho

Título

SPHEROID-BASED THREE-DIMENSIONAL CELL CULTURE ASSAY: A PROMISING ZIKA VIRUS STUDY MODEL

Introdução

In a tridimensional (3D) culture assay, cells are stimulated to grow in a dynamic environment mimicking in vivo conditions, enhancing proliferation and differentiation. Thus, such live systems can be considered important tools for studying virus-cell interaction and drug development. While 3D cell systems are widely used in distinct cancer research areas, in the field of virology these models are currently being developed and, therefore, yet to be proven suitable.

Objetivo (s)

This work aimed to standardize a spheroid-based 3D cell culture assay using Zika virus as a model, which can be further applied in virus replication and antiviral compounds screening studies.

Material e Métodos

The spheroids were produced from the Vero cell line, using the liquid overlay method. The viability of the spheroids was evaluated by Trypan Blue and the morphology was evaluated with the aid of the Fiji Software. A comparative quantification of ZIKV MR766 in Vero 2D and 3D cell cultures was performed using the endpoint assay and reducing the volume of spheroids (Fiji Software). 

Resultados e Conclusão

A total number of nine hundred and sixty spheroids within different days of growth were analyzed. An important volume increase of 40,3 % was observed over seven days. Spheroids formed from cells grown over 24 hours exhibited higher viability (100 %) than those from 48 hours (80%). Comparative quantification of ZIKV MR766 in both 2D and 3D cell cultures yielded similar results (p<0,05). Additionally, the reduction of spheroid volume at the infecting dose 50% (ID50), as determined by the endpoint assay, suggests their promising role in determining viral sample titers. Size measurements of infected and not infected spheroids can be used as a straightforward and easy method for Zika virus production and quantification.

Palavras Chave

3D cell culture; Zika Vírus; Vero Cells; Endpoint Determination