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Retroviral Mediated Gene Transduction: A Gene Transfer Technique to Deliver a Transgene in Cultured Cells Using Engineered Retroviral Vectors
Transcript
Collect fresh viral supernatant from the viral producer cell line. At this point, the cells should be well-spaced with a defined morphology. Next, carefully aspirate the top 2 milliliters of medium from each well of the 6-well plate containing the bone marrow cells. Avoid aspirating the bone marrow cells that are usually concentrated in the middle of the well.
Add 3 milliliters of freshly supplemented viral supernatant to each well of the 6-well plate containing bone marrow cells. After wrapping the plates in plastic, repeat the spin transduction, and then, place the unwrapped plates in the tissue culture incubator. After 24 hours on day three, remove and replace 2 milliliters of media from the wells.
