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Cellular characterization of epidermal growth factor-expanded free-floating neurospheres.

AuthorsLobo, María del Val Toledo; Alonso, F. J.; Redondo, C.; López-Toledano, M.A.; Caso, E.; Herranz, Antonio S.; Paíno, C.L.; Reimers, D.; Bazán, Eulalia
Keywordsprogenitor cells
growth factors
cell markers
adhesion molecules
stem cells
Issue Date2003
PublisherHistochemical Society
CitationJournal of Histochemistry and Cytochemistry 51: 89- 103 (2003)
AbstractNeural stem cells proliferate in liquid culture as cell clusters (neurospheres). This study was undertaken to characterize the epidermal growth factor (EGF)-expanded free-floating neurospheres derived from rat fetal striatum. We examined the ultrastructural and antigenic characteristics of these spheres. They consisted of two cell types, electron-dense and electron-lucent cells. Lucent cells were immunopositive to actin, vimentin, and nestin, whereas dense cells were immunopositive to actin, weakly positive to vimentin, and nestin-negative. Neurospheres contained healthy, apoptotic, and necrotic cells. Healthy cells were attached to each other by adherens junctions. They showed many pseudopodia and occasionally a single cilium. Sphere cells showed phagocytic capability because healthy cells phagocytosed the cell debris derived from dead cells in a particular process that involves the engulfment of dying cells by cell processes from healthy cells. Sphere cells showed a cytoplasmic and a nuclear pool of fibroblast growth factor (FGF) receptors. They expressed E- and N-cadherin, α- and β-catenin, EGF receptor, and a specific subset of FGF receptors. Because sphere cells expressed this factor in the absence of exogenous FGF-2, we propose that they are able to synthesize FGF-2.
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