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

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check this review for lentivirus.

[attachment=3580:LV_produ...ion_2006.pdf]

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Thanks for that, I think that should explain everything.

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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.

Cheers,


Lenti77

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