Retinal ganglion cells (RGCs) are central nervous system neurons with a very limited ability for axon regeneration. This unit details a cell culture technique, which can be used to functionally screen factors/compounds for their neuritogenic and neuroprotective effects on RGCs. In this protocol, the retina is isolated, digested in a papain solution, and after trituration, the RGCs are cultured. The neuritogenic effect of applied factors/compounds on RGCs in the medium is functionally determined by measuring the average neurite length of βIII-tubulin-positive RGCs in culture after 3 days. This protocol takes 3 to 7 days to perform depending on the application to complete, and is suitable to reliably test pharmacological and genetic approaches for their axon growth-promoting and neuroprotective potential on mature RGCs.
© 2010 by John Wiley & Sons, Inc.