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

Research output: Contribution to journalArticle

23 Citations (Scopus)

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 gamma-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 GABA(A) receptors, whereas activable kainate, alpha-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.
Original languageEnglish
Pages (from-to)1847-1856
Number of pages10
JournalCell Death and Differentiation
Volume15
Issue number12
Early online date12 Sep 2008
DOIs
Publication statusPublished - Dec 2008

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Neural Stem Cells
Lateral Ventricles
Neurons
Calbindins
Parvalbumins
alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid
Adult Stem Cells
GABA Receptors
Microtubule-Associated Proteins
Kainic Acid
Genetic Testing
N-Methylaspartate
Neuroglia
gamma-Aminobutyric Acid
Action Potentials
Cell Biology
Cell Survival
Potassium
Sodium
Calcium

Keywords

  • Action Potentials
  • Animals
  • Astrocytes
  • Cell Differentiation
  • Cell Division
  • Cell Line
  • Cell Proliferation
  • Cell Shape
  • Cerebral Ventricles
  • Ion Channel Gating
  • Mice
  • Neurons
  • Potassium Channels
  • Receptors, GABA
  • Receptors, Glutamate
  • Reproducibility of Results
  • Sodium Channels
  • Stem Cells
  • Time Factors
  • gamma-Aminobutyric Acid

Cite this

Setting the conditions for efficient, robust and reproducible generation of functionally active neurons from adult subventricular zone-derived neural stem cells. / Goffredo, D; Conti, L; Di Febo, F; Biella, G; Tosoni, A; Vago, G; Biunno, I; Moiana, A; Bolognini, D; Toselli, M; Cattaneo, E.

In: Cell Death and Differentiation, Vol. 15, No. 12, 12.2008, p. 1847-1856.

Research output: Contribution to journalArticle

Goffredo, D, Conti, L, Di Febo, F, Biella, G, Tosoni, A, Vago, G, Biunno, I, Moiana, A, Bolognini, D, Toselli, M & Cattaneo, E 2008, 'Setting the conditions for efficient, robust and reproducible generation of functionally active neurons from adult subventricular zone-derived neural stem cells', Cell Death and Differentiation, vol. 15, no. 12, pp. 1847-1856. https://doi.org/10.1038/cdd.2008.118
Goffredo, D ; Conti, L ; Di Febo, F ; Biella, G ; Tosoni, A ; Vago, G ; Biunno, I ; Moiana, A ; Bolognini, D ; Toselli, M ; Cattaneo, E. / Setting the conditions for efficient, robust and reproducible generation of functionally active neurons from adult subventricular zone-derived neural stem cells. In: Cell Death and Differentiation. 2008 ; Vol. 15, No. 12. pp. 1847-1856.
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AU - Biella, G

AU - Tosoni, A

AU - Vago, G

AU - Biunno, I

AU - Moiana, A

AU - Bolognini, D

AU - Toselli, M

AU - Cattaneo, E

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AB - 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 gamma-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 GABA(A) receptors, whereas activable kainate, alpha-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.

KW - Action Potentials

KW - Animals

KW - Astrocytes

KW - Cell Differentiation

KW - Cell Division

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KW - Cell Proliferation

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KW - Cerebral Ventricles

KW - Ion Channel Gating

KW - Mice

KW - Neurons

KW - Potassium Channels

KW - Receptors, GABA

KW - Receptors, Glutamate

KW - Reproducibility of Results

KW - Sodium Channels

KW - Stem Cells

KW - Time Factors

KW - gamma-Aminobutyric Acid

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JO - Cell Death and Differentiation

JF - Cell Death and Differentiation

SN - 1350-9047

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ER -