September 1994
Volume 35, Issue 10
Free
Articles  |   September 1994
Survival and differentiation of purified retinal ganglion cells in a chemically defined microenvironment.
Author Affiliations
  • J D Lindsey
    Department of Ophthalmology, Glaucoma Center and Research Laboratories, La Jolla, California 92093-0946.
  • R N Weinreb
    Department of Ophthalmology, Glaucoma Center and Research Laboratories, La Jolla, California 92093-0946.
Investigative Ophthalmology & Visual Science September 1994, Vol.35, 3640-3648. doi:
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      J D Lindsey, R N Weinreb; Survival and differentiation of purified retinal ganglion cells in a chemically defined microenvironment.. Invest. Ophthalmol. Vis. Sci. 1994;35(10):3640-3648.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

PURPOSE: To develop a fully defined medium and substratum that will support the survival and differentiation of purified retinal ganglion cells (RGCs) from newborn rats, and to evaluate beneficial effects of various hormones. METHODS: RGCs were purified from papain-dissociated retinal cells by a two-stage panning method and cultured in a fully defined medium on a substratum precoated with polyornithine and laminin. RGC purity was assessed by prelabeling RGCs with retrogradely transported horseradish peroxidase and by anti-Thy-1 immunocytochemistry. Various hormones were evaluated for their ability to promote RGC survival by adding them to the culture medium. RESULTS: Peroxidase labeling yielded purity estimates between 96% and 100%. Within 2 days, two distinct cell types are easily identified: a small cell characterized by thin, unbranched neurites and a large cell characterize by extensively branched neurites. Most small and large cells were round; however, some had elongated cell bodies. Both small and large cells express Thy-1 on their surfaces. Size-histogram analysis of all cells yielded a bimodal distribution. Survival was enhanced by seeding at higher densities and by the addition of hydrocortisone and progesterone to the cultures. CONCLUSIONS: This defined culture system should facilitate further studies on the direct mechanisms regulating retinal ganglion cell survival and differentiation.

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