FIG. 2.

Qualitative assessment of astrocyte reactivity at the interface with dissociated DRG cells. Astrocyte morphology and immunoreactivity was assessed using immunostaining and confocal microscopy for GFP (green), GFAP (red), and Hoechst (blue) at days 5, 10, and 15 in culture. Representative confocal projections are shown of astrocytes at the interface with dissociated DRG cells, and away from the interface, in an astrocyte only region at (A) low and (B) high magnification. Scale bar in (A)=150 μm and scale bar in (B)=40 μm. Over time, astrocytes in contact with/in close proximity to dissociated DRG cells become ramified and hypertrophic, which corresponds to an increase in staining for GFAP. GFAP, glial fibrillary acidic protein. GFP, green fluorescent protein. Color images available online at www.liebertonline.com/tec

Quantification of astrocyte reactivity at the interface with dissociated DRG cells: (C) GFP positive astrocytic cell nuclei (stained with Hoechst) were quantified at and away from the interface with dissociated DRG cells. Significantly more astrocytes were observed at the interface with dissociated DRG cells than away from the interface at day 5 and 10 in culture. (D) Quantification of GFP staining per cell revealed that astrocytes were becoming hypertrophic over time at the interface with dissociated DRG cells. No differences were observed in astrocyte size away from the interface. (E) Quantification of GFAP staining per cell revealed that astrocytes were becoming reactive at the interface with dissociated DRG cells, with significantly greater GFAP expression at day 15 in culture compared with away from the interface. *p<0.05, **p<0.01, ***p<0.001.