Hi,
My lab has stable clones from OSC2 cell line having puromycin resistance by lentiviral transduction. I would like to know if antibiotic should be added to the culture medium after the cells have been selected. some of them say, once they are selected, no antibiotic is necessary in the medium to maintain it. I would like to clarify. Experts please suggest
hi dear
please try to read this link hopefully it will be satisfactory
https://www.mirusbio.com/tech-resources/tips/generate-stable-cell-lines
Hi there,
If the cell line is stable, the selection marker is genetically stable (genomically inserted) therefore no further selection is absolutely required to maintain the cell line. But if you want to be sure about what you are actually culturing, antibiotic selection may help.
Yes, I agree with Dominique. Once selection is complete for lentiviral integrants, puromycin is no longer needed. Some people will tell you it helps prevent transgene silencing, but this isn't true. First, very low levels of resistance marker expression will yield a puro-resistance phenotype. Second, in dual promoter lentiviral cassettes, your GOI can be silenced independently of the puro selection cassette, so the puro resistance does not equate with transgene expression.
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