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Journal of Histochemistry and Cytochemistry, Vol. 51, 89-103, January 2003, Copyright © 2003, The Histochemical Society, Inc.

ARTICLE

Cellular Characterization of Epidermal Growth Factor-expanded Free-floating Neurospheres

Maria V.T. Lobob,c, F. Javier M. Alonsoa, Carolina Redondoa, Miguel A. López–Toledanoa, Enrique Casoc, Antonio S. Herranza, Carlos Luis Paínoa, Diana Reimersa, and Eulalia Bazána
a Departamento de Investigación, Hospital Ramón y Cajal
b Departamento de Biología Celular y Genética, Universidad de Alcalá
c Fundación FIO, Laboratorio de Oncología Molecular Aplicada, Servicio de Oncología Médica, Hospital Ramón y Cajal, Madrid, Spain

Correspondence to: Eulalia Bazán, Servicio de Neurobiología, Departamento de Investigación, Hospital Ramón y Cajal, Ctra. de Colmenar Km 9, 28034 Madrid, Spain. E-mail: eulalia.bazan@hrc.es

Neural 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, {alpha}- and ß-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.

(J Histochem Cytochem 51:89–103, 2003)

Key Words: stem cells, progenitor cells, ultrastructure, growth factors, cell markers, adhesion molecules, nestin


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