What cell type is used in this Zika virus infection protocol?

Human fetal brain neural stem cells are used as the target for Zika virus infection in this protocol.

How is the multiplicity of infection (MOI) determined and applied?

The MOI, representing the ratio of virus particles to cells, is set between 1 and 10 to control infection levels. The appropriate volume of Zika virus stock is mixed with the cell suspension accordingly.

What are the key steps to ensure only internalized virus remains with the cells?

After incubation, the mixture is centrifuged and the supernatant containing non-internalized virus is discarded. Cells are washed with DPBS and centrifuged again to further remove unbound virus.

How are the infected cells prepared for downstream analysis?

After infection and washing, cells are resuspended in culture medium and seeded onto extracellular matrix protein-coated multi-well plates for adhesion and further incubation.

What morphological changes are observed in infected neural stem cells?

Zika virus infection leads to significant alterations in cell morphology, cellular damage, and facilitates the spread of infection to neighboring cells.

Can this protocol be adapted for other types of analyses?

Yes, the infected cells prepared using this protocol can be used for immunocytochemical analysis and other virological or cellular assays.

Zika Virus Infection of Fetal Neural Stem Cells

JoVE Encyclopedia of Experiments

Neuroscience

0 views • 2:21 min • July 8th, 2025

$$\rightleftharpoonup{xx}$$ $$\longleftharp{xx}$$, $$\longrightharp{xx}$$,

Take a suspension of the Zika virus, a pathogenic virus, having the desired multiplicity of infection, the ratio of virus particles to cells.

Mix the virus suspension with human fetal brain neural stem cells.

Incubate the mixture, allowing the virus to bind to receptors on the cells, facilitating viral endocytosis.

Next, centrifuge the mixture and discard the supernatant containing non-internalized viral particles.

Resuspend the virus-infected cells in media and seed them into an extracellular matrix protein-coated multi-well plate, allowing cell adhesion.

Incubate the plate.

The internalized Zika virus utilizes the host cell machinery to produce new viral particles.

Viral replication significantly alters cell morphology, causing cellular damage and facilitating the spread of infection to neighboring cells.

The Zika virus-infected cells can be used for further analysis.

Resuspend the pellet in ZIKV stock at a Multiplicity Of Infection, or MOI, of between 1 and 10. The volume of ZIKV solution should not exceed 0.5 milliliters. Incubate the samples at 37 degrees Celsius for one hour. After, invert the tube two to three times, and centrifuge the mixture at 216 times g at room temperature for five minutes to obtain a cell pellet.

Remove the supernatant by aspiration. Then resuspend the cells with DPBS to rinse them. Next, centrifuge the mixture at 216 times g for five minutes, and remove the supernatant by aspiration. Resuspend the cells with 12 milliliters of the appropriate medium. Then load 500 microliters of the infected cells into a 24-well plate.