Advanced

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

Cacci, Emanuele LU ; Villa, A; Parmar, Malin LU ; Cavallaro, Maurizio LU ; Mandahl, Nils LU ; Lindvall, Olle LU ; Martinez-Serrano, A and Kokaia, Zaal LU (2007) In Experimental Cell Research 313(3). p.588-601
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... (More)
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 When growth factors were withdrawn, proliferation and expression of v-myc and telomerase were dramatically reduced, and the hc-NSCs differentiated into glia.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. (c) 2006 Elsevier Inc. All rights reserved. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
v-myc, immortalization, fetal, neural stem cells, cerebral cortex, telomerase, growth factor, differentiation, proliferation
in
Experimental Cell Research
volume
313
issue
3
pages
588 - 601
publisher
Academic Press
external identifiers
  • wos:000244283200014
  • scopus:33846850177
ISSN
1090-2422
DOI
10.1016/j.yexcr.2006.11.001
language
English
LU publication?
yes
id
7d4760e9-6888-4cfe-a006-20be7184a572 (old id 164137)
alternative location
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=17156776&dopt=Abstract
date added to LUP
2007-07-06 13:25:55
date last changed
2017-11-19 03:35:46
@article{7d4760e9-6888-4cfe-a006-20be7184a572,
  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 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. (c) 2006 Elsevier Inc. All rights reserved.},
  author       = {Cacci, Emanuele and Villa, A and Parmar, Malin and Cavallaro, Maurizio and Mandahl, Nils and Lindvall, Olle and Martinez-Serrano, A and Kokaia, Zaal},
  issn         = {1090-2422},
  keyword      = {v-myc,immortalization,fetal,neural stem cells,cerebral cortex,telomerase,growth factor,differentiation,proliferation},
  language     = {eng},
  number       = {3},
  pages        = {588--601},
  publisher    = {Academic Press},
  series       = {Experimental Cell Research},
  title        = {Generation of human cortical neurons from a new immortal fetal neural stem cell line.},
  url          = {http://dx.doi.org/10.1016/j.yexcr.2006.11.001},
  volume       = {313},
  year         = {2007},
}