Figure 4
Morphologic and immunocytochemical analysis of enteric neurospheres. (A) Appearance of neurospheres generated from neural stem cells (NSCs) isolated from embryonic day 12.5 intestine of pan-EGFP mice grown for 1, 7 and 21 days in culture. (B) Appearance of neurospheres on day 21 of culture showing green fluorescent protein (GFP) labeling of all cells, immunostaining for nestin (specific marker of neural stem cells), and the merged images. (C) Neurospheres were induced to differentiate into neurons and glial cells after withdrawal of growth factors, and were grown in poly-d-lysin/laminin-coated slide chambers for 4 days. Differentiated neurons and glial cells were recognized by Tuj1 and glial fibrillary acid protein (GFAP) staining respectively. D1, day 1; D7, day 7; D21, day 21. Scale bars: 50 μm.