Figure 5

Neural crest progenitor cells of the olfactory mucosa. a, b, Neural crest spheres cultured from single cells of the olfactory mucosa of P0-Cre/Floxed-EGFP (a) and Wnt1-Cre/Floxed-EGFP (b) mice in medium containing 1% methlylcellulose. c, RT-PCR analysis for the indicated neural crest markers from the following: E14.5 frontonasal tissue containing neural crest cells (frontonasal tissue), neurospheres prepared from E14.5 striatum of P0-Cre/Floxed-EGFP mice (CNS sphere), flow-cytometry-sorted GFP⁺ cells of the olfactory mucosa (OM primary), and cultured spheres from the olfactory mucosa (OM sphere) of postnatal 4 week P0-Cre/Floxed-EGFP mice. d, Neurons, glial cells, and myofibroblasts differentiated from clonal olfactory mucosa spheres, confirming the multipotency of the cells constituting these spheres. e, Cells from clonally cultured olfactory mucosa spheres were differentiated and stained for cell type markers: βIII-tubulin⁺ neurons, GFAP⁺ glial cells, and SMA⁺ myofibroblasts. The frequency of spheres that differentiated solely into neurons (N), glia cells (G), myofibroblasts (M) or into a combination of the three is presented as a percentage of the total number of spheres examined. Each bar represents the mean ± SD of three independent experiments, counting at least 20 spheres per experiment. f, Immunocytochemistry of cells differentiated from an olfactory mucosa sphere for OEC markers. Scale bar: (a) 50 μm, (d) 100 μm., (f) 100 μm.