hippocampal neuron transfection - (Aug/30/2005 )
As for hippocampal neurons transfection (18d old rat embryos neurons in culture), i currently use Lipofectamine 2000. The transfection rate is usually very low (from 2 to 10 transfected neurons out of 40-50000 seeded cells at the beginning).
I intend to try Fugene6 on these cultures, as it works perfectly well on cell lines such as COS-7 or even HEK-293.
Does someone have any experience of this ?
Several labs here at CSHL used gencarrier-2 for transfection of primary cortical neurons, PC12, C6..lines. We found this reagent gave us consistently high efficiency and very low cytotoxicity. Compared to lipo2000, fugene6, effectene, gencarrier-2 is better in all aspects.
I also used Lipfectamine and didn't get enough transfected Hippocampal neurons. Now I usually use Calcum precipitation and it's rate is better than Lipofectamine but not good enough,,
What is Generecin-2?
Gencarrier-2 is a lipoplex transfection reagent (new on market from epoch biolabs, not known to many people), apparently a mixture of cationic lipid, neutral lipid and others.... My colleague recently ordered one and tried on primary chick retinal neurons with ~30-40% efficiency and very low toxicity. Her excperience is that the younger cells the better the efficiency, and do it in serum-free, PBS-free and antibiotic-free medium.
Hi, Fair view:
I am thinking of trying GenCarrier 2 on retinal ganglion cells to deliver some siRNA. Do you know your colleague transfected siRNA or plasmid DNA? I am wondering whether I may ask for the protocol from your colleague. Such as what molar concentration she used, how long to leave it on, etc. Any information will be highly appreciated.