Transduction to Label PDX Tumor Cells: Introducing Lentivirus Expressing Fluorescent Marker into the Tumor Cell In Vitro

Lentivirus is a Retrovirus that contains RNA genetic material. This virus provides an efficient and stable method of introducing transgenes to cancer cell lines. Start by adding neuraminidase to a tube containing Lentivirus that expresses green fluorescent protein, or GFP. Keep the tube at 37 degrees Celsius for one hour to increase the virus's binding efficiency.

Next, centrifuge a tube containing tumor cell suspension to form a pellet. Add mammosphere media containing polybrene to resuspend the cells. Polybrene is a cationic polymer that increases Lentiviral infection efficiency. Now plate the optimal number of tumor cells per well in a six well tissue culture plate. Add the required amount of Lentivirus into each well of the culture plate.

Swirl the plate to mix the content, and keep at 37 degrees Celsius for 96 hours with a continuous supply of carbon dioxide. The Lentivirus enters the cell and releases RNA. The viral reverse transcriptase converts the RNA into double stranded DNA which integrates into the host genome. Place the plate under a fluorescent microscope and monitor cells for successful transduction. The transduced cells fluoresce due to GFP expression.

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