Tissue culture of adult human retinal ganglion cells.
OBJECTIVE
We wished to isolate and cultivate adult human retinal ganglion cells (RGC) from donor eyes.


METHODS
Small pieces of retina from donor eyes were plated in dishes and cultured with Ham's F12 medium with 10% serum for organ culture. For cell culture, cells were isolated by mechanical or enzymatic dissociation methods and cultured with F12 medium 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 cultures for which mechanical dissociation methods were used, the few cells that could be isolated showed poor viability. Better results were obtained with enzymatic dissociation methods. When cultured with medium supplemented with bFGF, some cells attached, spread, and sent out numerous dendrites, morphologically similar to RGC. These cells stained positively for neurofilaments and Thy-1 and negatively for glial fibrillary acidic protein (GFAP), indicating they were RGC.


CONCLUSIONS
Cell 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.
Hu
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Department of Pathology and Laboratory Medicine, New York Eye and Ear Infirmary, NY 10003, USA.
Ritch
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D000328:Adult; D002470:Cell Survival; D003470:Culture Media; D046508:Culture Techniques; D003712:Dendrites / Q000187:drug effects; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005904:Glial Fibrillary Acidic Protein; D006801:Humans; D007150:Immunohistochemistry; D008858:Microscopy, Phase-Contrast; D009414:Nerve Growth Factors / Q000494:pharmacology; D016900:Neurofilament Proteins; D012165:Retinal Ganglion Cells / Q000166:cytology* / Q000187:drug effects; D013936:Thymidine; D014019:Tissue Donors
D003470:Culture Media; D005904:Glial Fibrillary Acidic Protein; D009414:Nerve Growth Factors; D016900:Neurofilament Proteins; D016222:Fibroblast Growth Factor 2; D013936:Thymidine