Does anyone have experience with using Polybrene solution on neuroblastoma cells to improve lenti-viral transduction? I specifically use; BE2C, SY5Y, NBLS and LAN-5 cells. But any information on similar cells would also be good. I noticed that in other labs the use of Polybrene is part of a normal protocol for most cell types, but we don't use it. one paper said "Subconfluent SK-N-AS cells were grown in viral supernatant with 4μg/ml of polybrene (Sigma) for 48 hours followed by a 24 hour period in regular culture media." Origene website said its sometimes toxic for primary neuronal cells, so I am a bit cautious. Would you also recommend that it should be part of our normal protocol in general, HEK293s etc? I do get pretty good infection and it worked in the past for me, but it would be good to use less MOIs. Thank you in advance :)
I always use polybrene, I have not used the BE2C, SY5Y, NBLS and LAN-5 cells though. Usually I add 10 ug/ml polybrene while doing the infections and leave it for 24 hours. After what I remove the supernatant and add fresh complete medium, to allow the cells to recover from the infection procedure.
Patricia, just to clarify was that with neuroblastoma cells or another cell type? Thanks for answering before ! :)
Mostly cancer cell lines, but I have also used for primary endothelial cells (such as HUVEC and immortalized TIME cells) and normal human astrocytes (CC-2565). I hope this helps...
Hi, Jesica
we usually use polybrene for trasnduction of neuroblastoma cell line mainly N2A at the final conc 10 ug/ml and change the medium after 20 h to full medium for another 20 h followed by selection with antibiotic resistance mainly puromycin for 72 h at least. the efficiency of N2A transduction is from 40-60%
Hi Amr, thank you for you suggestions. What serum concentrations do you use?
the serum conc before transduction is 10% and after transduction i usually use 15-20% . another thing is that the confluency of cells before transduction had better not less than 50% as usually i use at least 60%
best wishes
In the SH-SY5Y cells I tried both 4 ug and 8 ug /mL both about doubled the transduction efficiency, with 8 ug being slightly better. I couldn't see signs of increased toxicity by eye.
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