They are in suspension and can be grown sera free, highly transfectable...
Since no one else has answered yet and I'm curious - why do you want to use cells in suspension as opposed to adherent cells?
My thought was it would be easier to scale up - big flasks etc, and pellet the cells to treat and put over gradients. I was thinking to just try and compare yield vs traditional method, but thought I would see if anyone has input .Thanks for query - thoughts on this?
I can definitely understand from the POV of scaling up...I did once visit a lab in Finland that was doing their AAV production in suspension cells but I think they were using baculovirus instead of plasmid transfection. Do you need large amounts of AAV for your experiments? Maybe a different serotype would be a less labor-intensive option?
Didnt want to go the Baculo route - we make several serotypes dependent on target/species and usually want 10e13 vgs at least. Using 20 - p150s at a shot seems cumbersome compared to a couple shaking flasks....in theory.....
Yup fair enough. Well all I can say it that I can't think of any reason why it WOULDN'T work, although the transfection reagent choice might be important. Good luck!
It may be late, but we have capability of doing large-scale production of AAV in 293 suspension cells by transient transfection in serum free medium. Although we have bioreactors, depending on the size of the Gene of Interest and your need to produce 1.0E+13 Vg, it is possible to do in couple of liters of cell culture in shake-flask.
Thanks Parminder, I also really appreciate your j Viro Meth pdf being available as elsevier is a pain. It is a nice study. I would like to try a bioreactor but they are a bit pricey, although i am looking at used ones. The amount of plasticware we go thru, using p150s, to get enough for a decent rat study seems crazy, in the long term. I have read there is shear stress/proper gas balance problems scaling up into large flasks (nonbioreactor). Thanks again
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