Hi everyone,
I’m currently performing retroviral transduction of primary T cells using viral supernatant generated from Plat-E (Platinum-E) packaging cells. I’m using Retronectin-coated plates to enhance transduction efficiency, but despite this, I’m observing very low transduction rates.
Here are some details of my protocol:
- Plat-E cells are transfected with the retroviral vector using a PEI max
- I collect viral supernatant at 48 hours post-transfection, filter it, and use it fresh.
- Viral supernatant is added to Retronectin-coated plates, followed by spinoculation (1500 × g for 1–2 hours at 32°C).
- I add activated T cells (CD4⁺) after spinoculation and incubate overnight.
- Despite this, my transduction efficiency remains low (
Dear Senthilkumar Thirumurugan
I would like to offer you a real efficient alternative to increase retroviral transduction of primary T cells with our LentiBlast Premium.
■ LentiBlast Premium is a novel patented chemical composition that dramatically increases lentiviral infection and transduction efficiency in any type of cells, adherent or in suspension, primary or cell lines. The properties of this reagent allow 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, LentiBlast Premium is non-toxic and totally compatible with cell viability.
LentiBlast Premium (LBP) was used in a variety of cells and demonstrated to be efficient in human T cells (Claireaux M., Nature Commun 2022 Jan 26;13(1):521 or Zheng L., Int J Mol Sci. 2017 Dec 20;18(12). pii: E2773 for example).
One of the advantage of LBP, beside its capacity to increase infection, is that it is totally non-toxic, so the doses can be increased to try finding the perfect conditions, and also it can be assocaited to retronectin protocol since its way of action is totally different.
I am sure it could be worth trying it,
If you need any information or if you want to try these reagents, please do not hesitate to contact me directly at:
best regards,
Cedric
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