Hi All,
I am doing the lenti-transduction of viral constructs in Neural progenitor cells, I require at least 2 wells of one six well plate lentivirus prepared from 293T to infect the one 12 well plate of NPCs. The construct was tagged with GFP. It takes more time to get the enough ceIls for my experiment. I am using lenti-x-concentrator to prepare the lentivirus.
I require to do more conditions to transduce the NPCs. Is there any way that I can minimize the preparation time or/and any other way to prepare the lentivirus.
You Suggestions would be helpful
Thank you
Dear Rubha Devi
I don't know if this post is still relevant,
In order to minimize the preparation or to raise the lentivirus titer without adding supplemental cost or effort, I would suggest using our LentiBlast Premium Transduction enhancer.
LentiBlast Premium is a patented modified poloxamer that we developed a couple of years ago and that acts by simultaneously neutralizing electrostatic repulsions between membrane and viral particles and enhancing viral fusion with cell membrane. Due to a favorable “membrane permeable effect” limiting the transmembrane potential changes, it is non-toxic and totally compatible with cell viability.
It is a real asset to viral infection because it allows to increase the effectiveness of the viral vector, thus reducing the MOI, and consequently the immune response of the cell. Secondarily, it reduces the cost of experience. And thus I believe it would be ideal in your experiment.
LentiBlast Premium was demonstrated to be efficient of multiple cell types from Hematopoietic Stem Cells, to T lymphocytes and I am sure it would be worth give it a try for your experiment.
Some examples of publications with LentiBlast Premium:
- HEK293: Brown AL. Sci Rep . 2021 Oct 25;11(1):21008.
- Bone marrow derived macrophages: Fonseca GJ. Nat Commun. 2019 Jan 24;10(1):414.
- Hematopoietic Stem Cells : Burton OT. Immunity. 2024 Jun 11:S1074-7613(24)00277-2.
- HMEC : Nassour J. Nature . 2023 Feb;614(7949):767-773.
- Myeloid progenitor : Verdys P. EMBO J. 2024 Jul 18. doi: 10.1038/s44318-024-00173-7.
- T lymphocytes CD4+ : Claireaux M. Nature Commun 2022 Jan 26;13(1):521.
- THP-1 : Chirayath TW. Nat Commun. 2024 Sep 18;15(1):8179.
you can have more infos at: https://ozbiosciences.com/lentivirus-retrovirus/113-418-lentiblast-premium-transduction-enhancer.html#/12-capacity-500_l
Would you like to give LentiBlast a try, please do not hesitate to contact me via ResearchGateor directly at: [email protected]; I will be glad to send you a sample !
best regards,
Cedric
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