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Title: Generation of human cortical neurons from a new immortal fetal neural stem cell line

Abstract

Isolation and expansion of neural stem cells (NSCs) of human origin are crucial for successful development of cell therapy approaches in neurodegenerative diseases. Different epigenetic and genetic immortalization strategies have been established for long-term maintenance and expansion of these cells in vitro. Here we report the generation of a new, clonal NSC (hc-NSC) line, derived from human fetal cortical tissue, based on v-myc immortalization. Using immunocytochemistry, we show that these cells retain the characteristics of NSCs after more than 50 passages. Under proliferation conditions, when supplemented with epidermal and basic fibroblast growth factors, the hc-NSCs expressed neural stem/progenitor cell markers like nestin, vimentin and Sox2. When growth factors were withdrawn, proliferation and expression of v-myc and telomerase were dramatically reduced, and the hc-NSCs differentiated into glia and neurons (mostly glutamatergic and GABAergic, as well as tyrosine hydroxylase-positive, presumably dopaminergic neurons). RT-PCR analysis showed that the hc-NSCs retained expression of Pax6, Emx2 and Neurogenin2, which are genes associated with regionalization and cell commitment in cortical precursors during brain development. Our data indicate that this hc-NSC line could be useful for exploring the potential of human NSCs to replace dead or damaged cortical cells in animal models of acute and chronic neurodegenerativemore » diseases. Taking advantage of its clonality and homogeneity, this cell line will also be a valuable experimental tool to study the regulatory role of intrinsic and extrinsic factors in human NSC biology.« less

Authors:
 [1];  [2];  [3];  [1];  [4];  [5];  [2];  [6]
  1. Laboratory of Neural Stem Cell Biology, Section of Restorative Neurology, Lund Strategic Research Center for Stem Cell Biology and Cell Therapy, BMC B10, Klinikgatan 26, University Hospital, SE-221 84 Lund (Sweden)
  2. Laboratory of Human Neural Stem Cell Research, Center of Molecular Biology Severo Ochoa, Lab CX-450, Autonomous University of Madrid, 28049 Madrid (Spain)
  3. Division of Neurobiology, Wallenberg Neuroscience Center, Lund University, BMC A11, SE-221 84 Lund (Sweden)
  4. Department of Laboratory Medicine, Section of Clinical Genetics, University Hospital, SE-221 85 Lund (Sweden)
  5. Laboratory of Neurogenesis and Cell Therapy, Section of Restorative Neurology, Wallenberg Neuroscience Center, Lund Strategic Research Center for Stem Cell Biology and Cell Therapy, University Hospital, SE-221 84 Lund (Sweden)
  6. Laboratory of Neural Stem Cell Biology, Section of Restorative Neurology, Lund Strategic Research Center for Stem Cell Biology and Cell Therapy, BMC B10, Klinikgatan 26, University Hospital, SE-221 84 Lund (Sweden). E-mail: Zaal.Kokaia@med.lu.se
Publication Date:
OSTI Identifier:
20972114
Resource Type:
Journal Article
Resource Relation:
Journal Name: Experimental Cell Research; Journal Volume: 313; Journal Issue: 3; Other Information: DOI: 10.1016/j.yexcr.2006.11.001; PII: S0014-4827(06)00459-9; Copyright (c) 2006 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; CELL PROLIFERATION; CEREBRAL CORTEX; FIBROBLASTS; GROWTH FACTORS; HYDROXYLASES; NERVE CELLS; NERVOUS SYSTEM DISEASES; POLYMERASE CHAIN REACTION; STEM CELLS; THERAPY; TYROSINE

Citation Formats

Cacci, E., Villa, A., Parmar, M., Cavallaro, M., Mandahl, N., Lindvall, O., Martinez-Serrano, A., and Kokaia, Z. Generation of human cortical neurons from a new immortal fetal neural stem cell line. United States: N. p., 2007. Web. doi:10.1016/j.yexcr.2006.11.001.
Cacci, E., Villa, A., Parmar, M., Cavallaro, M., Mandahl, N., Lindvall, O., Martinez-Serrano, A., & Kokaia, Z. Generation of human cortical neurons from a new immortal fetal neural stem cell line. United States. doi:10.1016/j.yexcr.2006.11.001.
Cacci, E., Villa, A., Parmar, M., Cavallaro, M., Mandahl, N., Lindvall, O., Martinez-Serrano, A., and Kokaia, Z. Thu . "Generation of human cortical neurons from a new immortal fetal neural stem cell line". United States. doi:10.1016/j.yexcr.2006.11.001.
@article{osti_20972114,
title = {Generation of human cortical neurons from a new immortal fetal neural stem cell line},
author = {Cacci, E. and Villa, A. and Parmar, M. and Cavallaro, M. and Mandahl, N. and Lindvall, O. and Martinez-Serrano, A. and Kokaia, Z.},
abstractNote = {Isolation and expansion of neural stem cells (NSCs) of human origin are crucial for successful development of cell therapy approaches in neurodegenerative diseases. Different epigenetic and genetic immortalization strategies have been established for long-term maintenance and expansion of these cells in vitro. Here we report the generation of a new, clonal NSC (hc-NSC) line, derived from human fetal cortical tissue, based on v-myc immortalization. Using immunocytochemistry, we show that these cells retain the characteristics of NSCs after more than 50 passages. Under proliferation conditions, when supplemented with epidermal and basic fibroblast growth factors, the hc-NSCs expressed neural stem/progenitor cell markers like nestin, vimentin and Sox2. When growth factors were withdrawn, proliferation and expression of v-myc and telomerase were dramatically reduced, and the hc-NSCs differentiated into glia and neurons (mostly glutamatergic and GABAergic, as well as tyrosine hydroxylase-positive, presumably dopaminergic neurons). RT-PCR analysis showed that the hc-NSCs retained expression of Pax6, Emx2 and Neurogenin2, which are genes associated with regionalization and cell commitment in cortical precursors during brain development. Our data indicate that this hc-NSC line could be useful for exploring the potential of human NSCs to replace dead or damaged cortical cells in animal models of acute and chronic neurodegenerative diseases. Taking advantage of its clonality and homogeneity, this cell line will also be a valuable experimental tool to study the regulatory role of intrinsic and extrinsic factors in human NSC biology.},
doi = {10.1016/j.yexcr.2006.11.001},
journal = {Experimental Cell Research},
number = 3,
volume = 313,
place = {United States},
year = {Thu Feb 01 00:00:00 EST 2007},
month = {Thu Feb 01 00:00:00 EST 2007}
}
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