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Yvonne Ou, Chay T. Kuo, Stuart J. McKinnon; Cell-based Therapy In Glaucoma: Co-cultures Of Neural Stem Cells With Glaucomatous Retinal Explants. Invest. Ophthalmol. Vis. Sci. 2011;52(14):2454.
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© ARVO (1962-2015); The Authors (2016-present)
To evaluate the neuroprotective potential of neural stem cell (NSC) transplantation in a murine glaucoma model using an ex vivo retinal explant organotypic tissue culture system.
Lineage-traced NSCs from the subventricular zone (SVZ) of the post-natal mouse brain were isolated and sorted by flow cytometry. Elevated intraocular pressure was induced by injecting hypertonic saline into the episcleral veins of wild-type adult mouse eyes. Retinal explants from glaucomatous and control eyes were co-cultured with lineage-traced NSCs and immunohistochemistry was performed.
After flow cytometry, lineage-traced SVZ-derived NSCs were differentiated into neurons and oligodendrocytes upon withdrawal of growth factors. Retinal explants derived from glaucomatous and control eyes were viable and expressed neuronal marker beta-III-tubulin, axonal marker ankyrin G, and astrocyte marker glial fibrillary acidic protein (GFAP). Co-cultures of lineage-traced NSCs with glaucomatous and control retinal explants demonstrated that these cells remain on the surface of the explants and did not migrate into the retina. Live imaging using two-photon microscopy of lineage-traced NSCs co-cultured with retinal explants derived from Thy 1-YFP mice did not show integration.
While lineage-traced NSCs co-cultured with retinal explants derived from glaucomatous and control mice did not show retinal integration, this ex vivo culture system is a powerful tool with which to study the potential of cell-based therapies in glaucoma.
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