TMEN508-16

[Rutcharin Potiwat]

1. What is the benefit of cell culture?
2. How to measurements the virus concentration using cell culture technique?

[Vanutsarin Khongsrisak]

1. What is the benefit of cell culture?
Cell culture is one of the techniques to study cell proliferation and understand their model system. This technique is widely used for investigating the cell under controlled conditions, generally outside of their neutral environment like when we needed to research virus biology, vaccine development, the role of genes in disease and health, and biopharmaceuticals. It’s used as a tool for many fields including;
– Models and systems in health and disease: study the route of interaction and infection route between cells and pathogens.
– Drug development and drug testing: Most of them are used to detect the efficacy or quality of drugs to find suitable chemicals compound before treating diseases, including screening novel chemicals and cosmetic products.
– Manufacturing of biopharmaceuticals: A large-scale production of antibiotics, enzymes, therapeutic monoclonal antibodies, therapeutic hormone formulations, etc.
– Other: Vaccine Production, Tissue Regeneration and Transplantation, Genetic Engineering, and Gene Therapy.

2. How to measure the virus concentration using the cell culture technique?
The measurement of virus concentration can be measured in two methods by using cell culture techniques, including plaque assay and TCID50 assay
Plaque assay
The plaque assay relies on determining the number of plaque-forming units (PFU) created in a monolayer of virus-infected cells. The assay requires the cultured susceptible cell to infect with the virus that interest. Bring the cultured cell that is fully growing on the well plate and add the serial dilution of the virus, incubated at 37oC in CO2 for around 3-4 days. After staining with 1% crystal violet in ethanol 3-5 drops, stand for a few minutes and wash the well plates to observe plaque formation. And calculate by using the equation to determine the virus concentration in mL.
50% Tissue Culture Infectious Dose (TCID50) assays, the use for the quantification of the virus. The process will be like the plaque by preparing serial dilutions of the virus and adding to the host cell, but they do not form the plaque. After the cell that contains the virus is incubated, we will calculate the percentage of dead cells from a cell viability assay by using a plate reader or observing under the microscope. The result showed a 50% infection dose per mL.

Vanutsarin Khongsrisak ID:6536456

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