FIG. 1.

Schematic of experimental strategy. (A) Gray matter cerebral cortical astrocytes were isolated from individual mouse pups at P1–P3. The astrocytes from a single pup are grown and purified in an individual flask and then used to compare the cellular responses to 2D and 3D environments. Each color represents an individual pup and cells obtained from it. Each experiment simultaneously compared astrocytes derived from three to four individual pups grown in 2D and 3D substrates and, in some cases, different medium formulations for each substrate. (B) Cells grown at a plating density of 7.5 × 10³ cells/cm² on a 2D substrate appear as flattened cobblestones and are immunoreactive for GFAP (red fluorophore). (C) Cells grown within the 3D substrate (7.5 × 10⁵ cells/ml) are distributed throughout the scaffold and are observed at multiple focal planes. The cells are immunoreactive for GFAP (red fluorophore) and appear much smaller than those in 2D (B). DAPI (blue) is used to stain cell nuclei. Scale bar: 50 μm. 2D, two-dimensional; 3D, three-dimensional; GFAP, glial fibrillary acidic protein. Color images available online at www.liebertpub.com/tea