Hi All,
Recently our lab want to delivery plasmids into primary cultured CNS neurons.I have tried lentivirus, lipo transfection before, and now we want to try electroporation. I checked online and didn't find any protocol for neurons. And we also want to delivery plasmids into neuron stem/progeniter cells, they were maintained as neuron sphere. But same thing, no protocol for neuron stem cell electroporation.
Did any one do it before? We were using Gene Pulser from bio-rad. And 293 cells working very good in this system.
Thanks!
Dear Monica,
As an alternative to "classic" transfection solutions that can be harmful for neurons, we have developped a transfection method that uses the Magnetofection*** technology.
NeuroMag, is the reagent of choice for neuron transfection. Initially developped for primary neurons, it was demonstrated by many to be highly efficient on any type of primary neurons (hippocampal, cortical, motor neurons, amygdal...) and also on neural stem cells.
you can find all the references mentionning the use of NeuroMag in our citation database: http://www.ozbiosciences.com/module/citationfinder/default
I am pretty sure NeuroMag will be of high importance for your transfection experiment, would you like to try it, please do not hesitate to contact me via researchgate or directly at : [email protected].
good luck with your experiment,
Cedric
MAGNETOFECTION
*** The Magnetofection technology uses a magnetic field to attract and concentrate complexes of magnetic nanoparticles and nucleic acid onto the cell surface as demonstrated by Grześkowiak BF et al, Pharm Res. 2014 Jul 18. (http://www.ncbi.nlm.nih.gov/pubmed/25033763).
Primary neurons have been successfully transfected with high efficiency and viability by electroporation using the MaxCyte STX. Users have developed screening assays for CNS applications using these biologically relevant transfected cells. See http://www.maxcyte.com/resources/Posters-Presentations/poster-Using-the-Right-Cell-for-the-Right-Application-SLAS2015.pdf for more details or contact MaxCyte at [email protected].
Dear Monica,
From your question, I assume that you are trying to electroporate your cells while they are still dissociated and in suspension, and then plate them or make them into neurospheres. If that is the case, you might want to see if someone at Wayne State has an Amaxa machine that you can try. It has electroporation protocols that are optimized for certain cell types, and a colleague of mine really likes it for neurons. The downside is that, as far as I know, you have to use their special buffers with their machine.
Good Luck!
Jill
- Badges
- Science method