Culture and characterization of mature primary retinal astrocytes.

<p><b>A)</b> A cartoon cross-section shows the restricted location of astrocytes in the adult eye (red lines). The inset fluorescent image corresponds to the ONH region stained for GFAP (red, arrows), with the dotted line indicating the site of detachment. <b>B)</b> Rapid dissociation and selective mature astrocyte culture conditions produced stellate, multi-processed cells that gradually transitioned to a polygonal morphology. A minority of cells displayed a distinct bipolar appearance characteristic of Müller glia (arrowheads). <b>C)</b> Cells were stained positively for astrocyte markers GFAP, vimentin, GS, Pax-2, and S100A. <b>D)</b> Cells did not stain for CD68, BRN3, or CD31 (green channel), but remained positive for GFAP (corresponding red channel merge below each panel). Nuclei are stained with DAPI (blue). (Scale bar = 50 µM, NFL; nerve fiber layer, GCL; ganglion cell layer, INL; inner nuclear layer, ONL; outer nuclear layer).