TISSUE-CULTURE OF ADULT HUMAN RETINAL GANGLION-CELLS

Purpose: We wished to isolate and cultivate adult human retinal gangli on cells (RGC) from donor eyes. Methods: Small pieces of retina from d onor eyes were plated in dishes and cultured with Ham's F12 medium wit h 10% serum for organ culture. For cell culture, cells were isolated b y mechanical or enzymatic dissociation methods and cultured with F12 m edium with 10% serum, with or without nerve growth factor (NGF) and/or basic fibroblast growth factor (bFGF). Results: In organ cultures, no neurite outgrowth from the retinal explants was observed. In cell cul tures for which mechanical dissociation methods were used, the few cel ls that could be isolated showed poor viability. Better results were o btained with enzymatic dissociation methods. When cultured with medium supplemented with bFGF, some cells attached, spread, and sent out num erous dendrites, morphologically similar to RGC, These cells stained p ositively for neurofilaments and Thy-1 and negatively for glial fibril lary acidic protein (GFAP), indicating they were RGC. Conclusions: Cel l cultures of human RGC can be established. This is a potential model system for studying effects of damaging and protective factors on RGC in vitro.