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A simple method for large-scale generation of dopamine neurons from human embryonic stem cells.

Morizane, Asuka LU ; Darsalia, Vladimer LU ; Guloglu, Oktar LU ; Hjalt, Tord LU ; Carta, Manolo LU ; Li, Jia-Yi LU and Brundin, Patrik LU (2010) In Journal of Neuroscience Research 88(16). p.3467-3478
Abstract
Dopamine (DA) neurons derived from human embryonic stem cells (hESCs) are potentially valuable in drug screening and as a possible source of donor tissue for transplantation in Parkinson's disease. However, existing culture protocols that promote the differentiation of DA neurons from hESCs are complex, involving multiple steps and having unreliable results between cultures. Here we report a simple and highly reproducible culture protocol that induces expandable DA neuron progenitors from hESCs in attached cultures. We found that the hESC-derived neuronal progenitors retain their full capacity to generate DA neurons after repeated passaging in the presence of basic fibroblast growth factor (bFGF) and medium conditioned with PA6 stromal... (More)
Dopamine (DA) neurons derived from human embryonic stem cells (hESCs) are potentially valuable in drug screening and as a possible source of donor tissue for transplantation in Parkinson's disease. However, existing culture protocols that promote the differentiation of DA neurons from hESCs are complex, involving multiple steps and having unreliable results between cultures. Here we report a simple and highly reproducible culture protocol that induces expandable DA neuron progenitors from hESCs in attached cultures. We found that the hESC-derived neuronal progenitors retain their full capacity to generate DA neurons after repeated passaging in the presence of basic fibroblast growth factor (bFGF) and medium conditioned with PA6 stromal cells. Using immunocytochemistry and RT-PCR, we found that the differentiated DA neurons exhibit a midbrain phenotype and express, e.g., Aldh1a, Ptx3, Nurr1, and Lmx1a. Using HPLC, we monitored their production of DA. We then demonstrated that the expanded progenitors are possible to cryopreserve without loosing the dopaminergic phenotype. With our protocol, we obtained large and homogeneous populations of dopaminergic progenitors and neurons. We conclude that our protocol can be used to generate human DA neurons suitable for the study of disease mechanisms, toxicology, drug screening, and intracerebral transplantation. © 2010 Wiley-Liss, Inc. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Neuroscience Research
volume
88
issue
16
pages
3467 - 3478
publisher
John Wiley & Sons
external identifiers
  • WOS:000284191200005
  • PMID:20981866
  • Scopus:78049437868
ISSN
1097-4547
DOI
10.1002/jnr.22515
language
English
LU publication?
yes
id
2fa3d994-2e1e-4c04-890d-435c057e2026 (old id 1710652)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/20981866?dopt=Abstract
date added to LUP
2010-11-05 14:18:15
date last changed
2016-10-13 04:31:18
@misc{2fa3d994-2e1e-4c04-890d-435c057e2026,
  abstract     = {Dopamine (DA) neurons derived from human embryonic stem cells (hESCs) are potentially valuable in drug screening and as a possible source of donor tissue for transplantation in Parkinson's disease. However, existing culture protocols that promote the differentiation of DA neurons from hESCs are complex, involving multiple steps and having unreliable results between cultures. Here we report a simple and highly reproducible culture protocol that induces expandable DA neuron progenitors from hESCs in attached cultures. We found that the hESC-derived neuronal progenitors retain their full capacity to generate DA neurons after repeated passaging in the presence of basic fibroblast growth factor (bFGF) and medium conditioned with PA6 stromal cells. Using immunocytochemistry and RT-PCR, we found that the differentiated DA neurons exhibit a midbrain phenotype and express, e.g., Aldh1a, Ptx3, Nurr1, and Lmx1a. Using HPLC, we monitored their production of DA. We then demonstrated that the expanded progenitors are possible to cryopreserve without loosing the dopaminergic phenotype. With our protocol, we obtained large and homogeneous populations of dopaminergic progenitors and neurons. We conclude that our protocol can be used to generate human DA neurons suitable for the study of disease mechanisms, toxicology, drug screening, and intracerebral transplantation. © 2010 Wiley-Liss, Inc.},
  author       = {Morizane, Asuka and Darsalia, Vladimer and Guloglu, Oktar and Hjalt, Tord and Carta, Manolo and Li, Jia-Yi and Brundin, Patrik},
  issn         = {1097-4547},
  language     = {eng},
  number       = {16},
  pages        = {3467--3478},
  publisher    = {ARRAY(0xb914c30)},
  series       = {Journal of Neuroscience Research},
  title        = {A simple method for large-scale generation of dopamine neurons from human embryonic stem cells.},
  url          = {http://dx.doi.org/10.1002/jnr.22515},
  volume       = {88},
  year         = {2010},
}