Recently , I am doing lentivirus-infection in HEK cells. According to the protocol of BROAD Institute: "https://portals.broadinstitute.org/gpp/public/resources/protocols", the infection always did in 6-well or 12-well plate.
I wonder if it is possible to infect cells in much bigger container, like T75 or T175 flask. That will be much convenient for large-scale stable cell making.
Some of you did the infection in big container? I ask myself, maybe, in smaller container with surface smaller than 10cm^2, the virus and cell distribution would be more uniform ?
Thank you for sharing any of your experience about this issue.
The smaller cell holder is easier to use higher the virus dose/concentration and easy to spin down virus particles to increase their interaction chances. Bigger flask still works in theory, but the efficiency should be foreseeing lower.
It depends on what type of protocol you are using. If you want incubate your virus plus cells overnight and you are working with adherent cells I would say to reduce as much as possible the media add a high number of cells, thus you will enhance the chances of cell and virus interaction.
However, if you plan on doing a few hours incubation of cells plus virus I would say add them into a 50ml Falcon tube and transfer to a T75 after infection for resting.
The last point, in case you are working with suspension cells, better leave them into a 50ml falcon tube either overnight or for a couple of hours (don't tight the lid all the way).
Be careful to avoid using too much pseudo/virus that could end up killing your cells,
Hope it helps.
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