Hi Nima Mohagheg! Regardless the viral vector type you choose for your transduction experiment, the most important thing is the purity and the concentration of viral vectors batch. So choose viral vectors with high titer (in TU/mL = transducing units) to obtain highly efficient CD14+ monocytes transduction. If you need more information check our website or contact us at [email protected]

Best regards, Nicolas

http://www.vectalys.com/

Hey!  Nima. You may hear that the type of vector is not important. Actually it is. Lentiviral Vectors work better with cells that are not dividing opposite to the Retrovirus wich are more efficient with dividing cells. Also there are two ways to achieve high titers of suspension cells (we all know how hard that is!) you must get your hands on some Retronectin (fibronectin) or other matrix of some sort, this will allow  the suspension cells to attach to the plate and easily bud with the lentivirus.  If you have a plate rotor use it, centrifugation for 1 or 2 hours at t 1,000 - 2,000Xg will help suspension cells to enter in contact with the virus at the bottom. Even if you concentrate your lentivirus you need stimulation of some sort for monocytes and other suspension cells like T cells. Hope this will help you. Good Luck!

Nicolas and German; thanks for the replies.

I mostly wonder if various cell lines (not merely dividing vs non-dividing) are more susceptible to uptake certain kind of viruses over others. And how could we find out  about that. Does people just try and error? Or there are studies focused on this aspect of different viral-vectors.

Trial and error, pretty much! The type of virus and also the promoter choice will be important. Will your cells stay in culture and divide or will you harvest them soon after infection? If they stay in culture for a while then AAV will be no good as it doesn't integrate. Lentivirus will almost certainly be your best bet, it works well in T-cells but I believe they have to be stimulated first. I use a EF1a-GFP in T-cells and the transduction efficiency is very good provided you use a high enough MOI.

 I think that your that the answer you seek is more complex than you think.  If you want to choose a viral vector that offers you high titers you must choose Amphotropic Lentiviral vectors like  pCMV-Gag-Pol vector as Capsid and Envelope vector, either pCMV-VSVG, pCMV-Ampho, or pCMV-Eco or a similar system. But tropism depends not only of the type of virus but also the receptor of the cell, and even PH. In the case of CD14 monocytes CMV entry could use CD14 receptor and TLR-4. So next time you want to infect cells check for tropism and cell receptors, there are a lot of informative papers about it http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3144558/

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1368960/