Primary Retinal Cultures as a Tool for Modeling Diabetic Retinopathy: An Overview
Morphologic and immunocytological characterization of dissociated ((a)–(h)) and organotypic ((i), (j)) primary rat retinal cultures. In dissociated mixed cultures, the heterogeneous cell population is composed of differentiated neurons, labeled for axonal (Tau, (a)) or dendritic (MAP2, (b)) microtubule-associated proteins and synaptophysin (Syn) (b) and of Müller cells, positive for vimentin (Vim) (c). In the neuronal population, photoreceptors form rosette-like aggregations (SEM, (d)), positive for rhodopsin (e). The positivity for the inhibitory neurotransmitter GABA indicates the presence of amacrine or horizontal cells (e). Thy1.1-immunolabeled ganglion cells are also present ((f), inset). Coimmunostaining of vimentin, a marker of Müller cell cytoskeleton, and NeuN, which labels neuronal nuclei, proves the mixed neuronal/glial composition of the cell culture (f). Following special culturing protocols, retinal cell cultures can be enriched in neurons (g) or in Müller cells, as shown by the specific marker S100B (h). In organotypic cultures, retinal tissue structure in preserved. In (j), Müller glia projections, positive for pERK1/2 after short-term treatment of HG, cross the inner plexiform layer and expand in the ganglion cell layer. (i) Control sections. Bar = 10 m.