Hi everyone! I work with NK-92 cell line (ATCC) and am trying to transduce them with a CAR. After several attempts of transduction, I’ve achieved a place where I feel hopeless. I’ve tried to alter many different variables in the protocol, since at the virus production stage - using HEK 293-T cells and lipofectamine 3000 transfection protocol - till enhancements in the transduction protocol. I concentrated the virus, used Vectofusin and spinoculation to improve transduction, tested my protocols on other cell types and they’re working. I am now producing the viral particles with a vector for the envelope that is supposed to be more effective on NK cells that the commonly used VSVG, BaEV-LV, and I‘ve also tried the same protocol that the authors present (Bari R et al, 2023; doi: 10.3389/fimmu.2019.02001), but apart from one attempt (in which the cells stopped proliferating after transduction and slowly died, and which I could not replicate, even doing exactly the same steps), I still didn’t get an efficiency rate greater than 3%. I don’t know if there’s a tip or any determinant step in the protocol that I’m missing, but any kind of help would be very welcome!
- Badges
- Science topic
- Similar topics
- Biological Science
- Microbiology
- Virology
- Virus