We need a high titre of active virus to be used in the bioassay. But the bank seems to be losing its infectivity. Even upon several passages (Medium: DMEM - 2% FBS) the desired titre is not yet achievable.
I doubt whether the medium with 2% FBS supports the proper health of the cells and hence the viral replication. So I would suggest to go for basic medium and also use polybrene, which enhances the infectivity of the virus. I hope this helps. Good luck
What is your current titer and what titer do you want to achieve? There are several things that you could try:
1. optimize infection by inoculating cells with various MOIs. Suggest to start with MOI of 0.1, 0.01, etc.
2. Concentrate the virus stocks.
The tire required is to give EC50 for a fixed drug dose and cell density (20 x 10^4 cells/ml respectively). I have been trying to concentrate the stock, but now as the no. of passage increases, the cell lysis is decreasing. It seems that the virus is losing infectivity in the cell line. When I employed the same titre on different cell line (same no. of cells) it exhibited 100% lysis.
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