I'm currently trying to transduce the rADSC mentioned above using several lentiviruses (containing different genes and the same promoter, 1: Zsgreen fluorescent protein/other+ puromycin resistance with EF1a promoter). I'm using Lenti-X Accelerator by Takara magnetic beads system, instead of the classical polybrene method to reduce toxicity.

Previously I had successfully infected other cell lines - rat adenocarcinoma of breast (Rba) cells and human embryonic kidney (HEK) cells with a wide range of multiplicities of infection: from 1 to 100. Both transduced Rba and HEK cells survived puromycin selection, and cells transduced with ZsGreen protein became fluorescent.

I'm wondering what may be the reason for no transduction in rADSC? Cells seem healthy after transduction, however soon after the puromycin selection starts they all die.

The manual for Lenti-X Accelerator gives information about using a penicillin/streptomycin-free medium during transduction - I did use an antibiotic-free medium for both HEK and Rba cells transduction and AA+ for rADSC transduction, but is it possible that this may be the reason for 0% transduction efficiency?

Thank your for your insight