Methods for Culturing Primary Adult CNS Neurons

In 1910, Harrison published the first report of frog embryonic sympathetic ganglia grown in hanging drops of lymph for a few days, where single neurons extended nerve fibers with complex growth cones extending and contracting over time towards target cells. The revolutionary ability to culture embryonic and post-natal neurons for extended time periods demonstrated that developing neurons could form plexi of interconnected nerve fibers. However, over the years it has proven to be exceedingly difficult to consistently culture adult central nervous system (CNS) neurons; while several instances of success in culturing fewer than 10 neurons have been reported, a major unresolved methodological challenge in neuroscience has been the ability to culture adult CNS neurons from any region consistently and in tractable numbers. It has been possible to culture adult dorsal root ganglia neurons for reasons that remain unclear; these neurons also retain an ability to regenerate throughout life. The value of CNS neurons in advancing understanding of intact adult neuronal function, response to injury, and as a screening system for candidate therapeutics is clear. Here we report for the first time the ability to consistently culture adult neurons of specified phenotype in large numbers over extended time periods. We further provide evidence for the utility of these cultures in advancing studies of CNS injury.

Learning Objectives:

1. Successful culture of adult primary neurons from the central nervous system

2. Electrical recordings and stimulation of neural networks in vitro

3. Practical applications for primary adult CNS cultures