Tissue culture of adult human retinal ganglion cells

Dan Ning Hu, Robert Ritch

Research output: Contribution to journalArticlepeer-review

34 Scopus citations

Abstract

Purpose: 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.

Original languageEnglish
Pages (from-to)37-43
Number of pages7
JournalJournal of Glaucoma
Volume6
Issue number1
DOIs
StatePublished - Feb 1997
Externally publishedYes

Keywords

  • Basic fibroblast growth factor
  • Cell culture
  • Nerve growth factor
  • Organ culture
  • Retinal ganglion cells

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