Hello,
I am experiencing problems with expressing GOI2 in GOI1-IRES-GFP:GOI2 cassette in CD4/CD8 cells separated from PBMCs using Miltenyi beads.
However expression of the same GOI2 (using the same construct) seems to be ok in Jurkats.
I am trying to figure out what's the big difference between this two cell lines - what can be the issue?
Would be happy to receive any suggestion/explanation, because for me Jurkats and T-cells are quite similar and they are always sued as a lymphocytes cell model due to their "easiness".
Thank you in advance!
Hello,
in my experience there are huge differences between primary T cells and established cell lines regarding transduction efficiencies. We transduced both Jurkats and primary CD4 / CD8 T cells with Lentiviral supernatants and we observed that primary T cells are much more resistant. Maybe you could try to concentrate your supernatant in order to reach a higher MOI.
- Badges
- Science method