Figure 1.

Isolation of endogenous precursor cells from the adult mouse dentate gyrus. A; Coronal 300 µm vibratome section through the brain of a Nestin-GFP transgenic mouse. B; As a second step, the dentate gyrus was isolated from the rest of the tissue by microdissection. The dissected dentate gyrus is shown at the top. C; The dissected dentate gyrus, as depicted in B, was further trimmed by placing two cuts that separate the dentate gyrus from ventricular tissue (of the 3^rd ventricle) and area CA3 (see also D). Nestin-GFP expression in the subgranular zone is seen with high concentration at the medial tip of the granule cell layer. D; Schematic drawing depicting the dissected region that was used for isolation of the precursor cells. The localization of the potentially contaminating ventricular areas is highlighted in red. The dashed lines indicate the cuts described in C. E; The tissue was homogenized and separated on a Percoll density gradient. Proliferating precursors were present in the region with density>1.053 gm/ml (>22% Percoll). The Erythrocytes were enriched in the region with density>1.075 gm/ml (>65% Percoll). F; phase contrast image of mouse precursor cells grown as adherent cultures. Note the phase bright appearance that is characteristic of proliferating precursor cells. G; Incubation with BrdU (20 µM) revealed that isolated nestin-expressing cells were proliferating. Nestin (Green), BrdU (Red).