Although new culture conditions enable homogeneous and long-term propagation of radial glia-like neural stem (NS) cells in monolayer and serum-free conditions, the efficiency of the conversion of NS cells into terminally differentiated, functionally mature neurons is relatively limited and poorly characterized. We demonstrate that NS cells derived from adult mouse subventricular zone robustly develop properties of mature neurons when exposed to an optimized neuronal differentiation protocol. A high degree of cell viability was preserved. At 22 days in vitro, most cells (65%) were microtubule-associated protein 2+ and coexpressed -aminobutyric acid (GABA), GAD67, calbindin and parvalbumin. Nearly all neurons exhibited sodium, potassium and calcium currents, and 70% of them fired action potentials. These neurons expressed functional GABAA receptors, whereas activable kainate, -amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid and N-methyl-d-aspartic acid receptors were present in approximately 80, 30 and 2% of cells, respectively. Antigenic and functional properties were efficiently and reliably reproduced across experiments and cell passages (up to 68). This is the first report showing a consistent and reproducible generation of large amounts of neurons from long-term passaged adult neural stem cells. Remarkably, the neuronal progeny carries a defined set of antigenic, biochemical and functional characteristics that make this system suitable for studies of NS cell biology as well as for genetic and chemical screenings

Setting the conditions for efficient, robust and reproducible generation of functionally active neurons from adult subventricular zone-derived neural stem cells / D. Goffredo, L. Conti, F. Di Febo, G. Biella, A. Tosoni, G. Vago, I. Biunno, A. Moiana, D. Bolognini, M. Toselli, E. Cattaneo. - In: CELL DEATH AND DIFFERENTIATION. - ISSN 1350-9047. - 15:12(2008 Dec), pp. 1847-1856.

Setting the conditions for efficient, robust and reproducible generation of functionally active neurons from adult subventricular zone-derived neural stem cells

D. Goffredo
Primo
;
L. Conti
Secondo
;
G. Vago;A. Moiana;E. Cattaneo
Ultimo
2008

Abstract

Although new culture conditions enable homogeneous and long-term propagation of radial glia-like neural stem (NS) cells in monolayer and serum-free conditions, the efficiency of the conversion of NS cells into terminally differentiated, functionally mature neurons is relatively limited and poorly characterized. We demonstrate that NS cells derived from adult mouse subventricular zone robustly develop properties of mature neurons when exposed to an optimized neuronal differentiation protocol. A high degree of cell viability was preserved. At 22 days in vitro, most cells (65%) were microtubule-associated protein 2+ and coexpressed -aminobutyric acid (GABA), GAD67, calbindin and parvalbumin. Nearly all neurons exhibited sodium, potassium and calcium currents, and 70% of them fired action potentials. These neurons expressed functional GABAA receptors, whereas activable kainate, -amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid and N-methyl-d-aspartic acid receptors were present in approximately 80, 30 and 2% of cells, respectively. Antigenic and functional properties were efficiently and reliably reproduced across experiments and cell passages (up to 68). This is the first report showing a consistent and reproducible generation of large amounts of neurons from long-term passaged adult neural stem cells. Remarkably, the neuronal progeny carries a defined set of antigenic, biochemical and functional characteristics that make this system suitable for studies of NS cell biology as well as for genetic and chemical screenings
neural differentiation ; neural stem cell ; adult stem cells ; in vitro differentiation ; neurones
Settore MED/08 - Anatomia Patologica
Settore BIO/14 - Farmacologia
dic-2008
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/60475
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