Hi all,
Looking to see if anyone has sorted neurons from SC tissue using FACS before? Interested in trying this, will likely follow Ernfors 2018 paper, but wanted to see if anyone else has any good protocols out there?
Thanks!
The reason you don’t see it much is because getting intact neurons from adult spinal cord is very challenging. The Haring paper is an exception. They used 3-4 week old mice and used transgenic lines to enrich for the neurons. You’d probably need to do the same.
Dissociating adult spinal cord gets plenty of glia, but the neurons just don’t seem to want to come out, and if they do, they’re not super viable.
Try it. See how you do. I’d use the protocol from Haring 2018 as a starting point. What is your application? RNA-seq?
You might consider using nuclei instead of cells. Much easier to obtain and you can sort on NeuN or use a transgenic line