Hello,
After we co-transfected our hek293 packaging cells with RC, pHelper, GOI for AAV production; can we stock some of them for future experiments with using freezing and thawing method? Are viral particles become dysfunctional or being degraded with freezing&thawing? Im asking that for just theoritical background and for eliminating purification steps for vectors from bacterial stock plates. If there is no shortcut solution for eliminating first purification from agar plates, I think, keeping stock as untransfected format at bacterial glycerol stock is more efficient and unproblematic. ( I asked because i saw large manufacturing systems use producer cell lines (with LTR transduced lines for proviral integration)
Thank you in advance for the answers.
Hi, I am not quite sure what you mean by "future experiments". In case you mean using transiently transfected HEK cells for AAV production on the long term, like if it was a producer cell line, then the answer is no, it is not possible, because the transfection is transient. Producer cell lines are a different story.
But it is certainly possible to collect the transfected and AAV producing HEK cells and freeze them for storage, if that is what you meant, the AAV particles should be fairly stable in this form. By doing this, you only delay the time of AAV purification a bit for example until you add more transfected cells to the stock. For doing actual experiments, the purification of AAV particles will have to be finished anyways. And again, these transiently transfected cells are for one time use.