Characterization of neurospheres converted from rat ADSC. A: Under phase contrast, neurospheres grow as spheres of floating cells. B, C: Immunocytochemistry of neurospheres. Neurospheres were triturated before being examined. About 85 ± 7% of the neurosphere cells express nestin (B), whereas only a small part of neurosphere cells express fibronectin (C). D-G: Neurosphere cells can differentiate into neural cells. Neurospheres were cultured and maintained for 10 days in Neurobasal® medium supplemented only with B27 supplement on poly-L-lysine-coated substrate. Neurospheres attach to the bottom of the culture dish and protrude cell processes (D), some cells gain a neuronal morphology and the processes grow much longer 10 days after plating (E); Immunocytochemistry shows that part of the differentiated neurosphere cells express neuronal marker beta-tubulin III (F) and glial marker GFAP (G) 10 days after plating. Nuclei are labeled with DAPI (blue). Bar, 50 μm.