Do more people use cDNA, siRNA, mRNA or miRNA? Would love to hear what people are using?
We have used retrovirus to infect dental pulp stem cells. We have not tried neurons derived from DPSC yet. Retrovirus works well. What are you trying to do? Transient knock down?
We as many others use lentivirus and AAV mediated viral transduction, with all of the above cell types. We have used magnetic transfection beads and electroporation as well with some succsess. Neural stem cells should transfect quite easily with any of the above but long term expression during their subsequent differentiation may be difficult. I know of people who are using lentiviral mediated ShRNA delivery in neurons with good results
Thanks everyone for your responses! Keep them coming! I'm looking to find out what researchers prefer when transfecting primary neurons, astrocytes, IPS derived neurons, neural stem cells. Also, what is most important: high transfection efficiency, low variability, low toxicity etc.?
Primary neurons, astrocytes and IPSC derived neurons are non-dividing cells. As such, you want to keep efficiency high and toxicity low. Variability is something you should keep out of all of your experiments if possible.
As for the neural stem cells, we typically infect stem cells prior to neuronal differentiation - when they are still dividing like fibroblasts - and then perform selection for cells with the construct we want inserted. This is the best way to get things into neurons is to get them to stably integrate into the precursors BEFORE you make the neurons.
Hello! For primary hippocampal and cortical neurons, I usually do nucleofection before plating, with Amaxa kit. High efficiency and low damage.
For transfection after plating, I use Lipo200. It works well and it's not toxic.
Dear Nick,
Does your mentioned product is compatible with mRNA upto 1.5 kbp and in iPS cells as well as neurons?
Thanks a lot!
Best regards,
Ashutosh
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