I would like to deliver some reporter plasmids into neural rosettes without lifting them from the coating. I was wondering if anyone has a protocol for doing so either by viruses or by lipofection or any similar methods.
Dear Navneet,
There is an alternative to classic transfection that gives real good results with preserving high viability in hard-to-transfect and primary cells: Magnetofection (the use of magnetic field to concentrate magnetic complexes of vector onto the cell surface).
Beside the fact that Magnetofection is ideal for any primary cells, this method was demonstrated in 2007 by Buerli et al (Nature Protocol) to efficiently transfect primary neurons.
Since then, numerous paper showed its efficiency for any neuron types (from neural stem cells, to astrocytes) and a specific magnetofection reagent dedicated to neurons was developped : NeuroMag. For example NeuroMag was used by Charrier et al, in Cell, 2012 to transfect mouse primary hippocampal neurons at DIV 18-21! (http://www.ncbi.nlm.nih.gov/pubmed/22559944)
or more recentlyby Courchet et al, in Cell, 2013 to transfect cortical neurons. (http://www.ncbi.nlm.nih.gov/pubmed/23791179)
for more information, you can have a look at :
http://www.ozbiosciences.com/primary-neurons-transfection.html
I'm pretty sure this reagent will give you reliable efficiency, you can contact me directly at [email protected] if you need more informations or if you want to try Magnetofection.
Good luck for your experiments,
Cedric
Hi Cédric,
Thanks for your reply. The NeuroMag does indeed sound interesting but I was wondering whether it could be specifically adapted for progenitor cells in neural rosettes as I've already tried lipofectamine methods.
Any information in this regard would be very helpful.
Thanks!
Hi Navneet,
indeed, NeuroMag was used in Neural Stem Cells ; in collaboration with C. Beclin from the french national center of scientific research (CNRS) we've published a paper in Biotechniques (2011) demonstrating NeuroMag efficiency in Neural Stem cells without affecting their differenciation potential.
I'm pretty sure that it can be adapted for your progenitor cell type.
I would be glad to send you samples to test ;
Hope this will be usefull,
have a nice day,
Regards,
Cedric
Article High transfection efficiency of neural stem cells with magnetofection
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