Tissue Culture Techniques-2 and CPE

Virus Infection and Cell Culture

• Applying three cycles of freezing and thawing on infected cells helps to release the viral particles from the cells, allowing for efficient viral purification and concentration.

Virus Preservation

• Preserving the supernatant fluid containing the virus in an ultra-deep freezer is important to prevent viral degradation and inactivation, ensuring the virus remains viable for further experiments.

Virus Infection of Adherent Cells

• Incubating the vessels at 37ºC for 1-2 hours with gentle tilting every 10 minutes after adding the virus suspension to adherent cells facilitates the attachment and penetration of the virus into the cells, promoting efficient infection.

Cell Monolayer Preparation

• Washing the cell monolayer with sterile PBS after removing the inoculum from adherent cells is necessary to remove any residual virus and prevent further infection, ensuring a clean environment for cell growth and observation.

Monitoring Cytopathic Effects

• Daily observation until the appearance of complete Cytopathic Effects (CPE) in suspended cells is significant because it allows for the detection of viral replication and the assessment of the infectivity of the virus, enabling researchers to monitor the progression of the infection.