I want to transduct bone marrow derived mice dendritic cells (day 0 and then day 5, and day 5 only) with lentivirus containing my gene and GFP.
I got low transduction rate
High level of DCs death
I don’t have GFP expression after 3 days (transduction of HEK cells in the same time result more than 80% GFP expression)
(For a comparison, transfection of DCs with FITC conjugated siRNA with lipofectamin result about 60% transfection and FITC expression in flowcytometery)
What is the problem?
Which point is critical?
Is there any similar experience with GFP expression and how the protocol is?
Any comment please?
Thank you
Dendritic cells are difficult to transfect with lentiviruses, lipofectamine, electroporation etc.
The best method in our lab was the Amaxa kit, which could transfect several genes better. Moreover, DCs are very sensitive and die a lot after transfection.
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