Protocol Online logo
Top : Forum Archives: : Transfection and Transduction

recommendations for a viral titer cell line - (Sep/12/2007 )

Greetings earthlings,

I'm working my way through the Invitrogen lentiviral expression system and finally I'm at the point of making some virus. After which I want to carry out a viral titer- which all seems straight forward. They're recommending the 'gold standard' HT1080 cell line for determining titer, which is a fibrosarcoma cell line.However, they also say that any cell line is okay (diving and non-primary). Doing a quick search- it seems that anything is almost acceptable- HeLa, cells etc etc. Unfortunately the cells that I'm growing at the moment fit into the primary (pain in the butt to culture)category thus rendering them useless for titering.
Before I fork out more grant money for yet another cell line. I wanted to ask what all you lentiviral experts would recommend as the best viral titer cell line? Is it better to have a cell line that is most similar to your final cell type for example an epithelial cell line for epithelial cell types?

Cheers in advance
Any advice, as usual, much appreciated smile.gif

-lost in the lab-

check this review for lentivirus.



QUOTE (scolix @ Sep 14 2007, 07:08 AM)
check this review for lentivirus.


Thanks for that, I think that should explain everything. smile.gif

-lost in the lab-

I'm using the same kit (BLOCKiT miRNA lenti's though), I have titered on HT1080's, and they are susceptable to transduction but the resultant GFP is dim. I did a side by side comparison with 293T cells, and found that they are more sensitive to transduction and thus would yield a more accurate viral titer than HT1080's.

293T's are relatively common and you could possible get a flask from someone. If not you can order them from ATCC for about 120 USD.