I have been trying to use lentiviral on primary culture to knockdown genes. My protocol says that try to use serum free media to resuspend concentrated virus because serum inhibits infection. I culture my cells in DMEM+10%FBS and I have tried to substitute this with serum-free media. It gives a reasonably good transfection rate but the substitution makes the cells behave a bit weird.
Since I am knocking down genes I am trying to get close to 100% infection rate. I am using a very high virus concentration and I use polybrene. So I don't think I can do much on those aspects. I am just wondering how much serum could inhibit virus infection-- true I could just do an experiment to see, but virus is precious and so are the cells.... So I am wondering if any of you have ever tried infecting PRIMARY CULTURE with lentivirus in serum-containing media. Cell lines do not count since there are much easier to infect.
The culture I am doing is radial glia culture. I dissect P0 SVZ from mice and culture the cells until confluency in DMEM+10%FBS. After confluency I switch to 2% FBS for differentiation. I infect during proliferation stage so that I could spare a few days for the gene to knockdown at differentiation. So if you also do this culture and know any substitution serum-free media please also let me know.
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