Neonatal retinal ganglion cell cultures of high purity: effect of superior colliculus on their survival.
A method is described for obtaining retinal ganglion cell (RGC) cultures of high purity. RGC were retrogradely labelled in vivo with either the fluorescent dye True Blue and horseradish peroxidase (HRP), or with FITC-conjugated HRP. Following dissociation, RGC were separated from the intrinsic cells of the retina using fluorescent activated cell sorters, on the basis of their greater size and fluorescent intensity. When the sorted cells were cultured, RGC could be subsequently identified by their HRP labelling. Using such criteria, cultures in which 75% of the cell population consisted of RGC could be regularly obtained. This represents a 150-fold increase in RGC concentration over whole retinal cultures. The survival of the sorted RGC could be greatly enhanced when they were cocultured with their target tissue, the superior colliculus.
D000818:Animals; D000831:Animals, Newborn / Q000033:anatomy & histology* / Q000502:physiology; D002469:Cell Separation; D002470:Cell Survival; D002478:Cells, Cultured; D008722:Methods; D051381:Rats; D012160:Retina / Q000166:cytology*; D012165:Retinal Ganglion Cells / Q000166:cytology* / Q000502:physiology; D013477:Superior Colliculi / Q000502:physiology*