| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
EMBRYONIC STEM CELLS |
1 Udall Parkinson's Disease Research Center for Excellence; Center for Neuroregeneration Research; McLean Hospital/Harvard Medical School, Belmont, MA
2 Udall Parkinson's Disease Research Center for Excellence; Molecular Neurobiology Laboratories; McLean Hospital/Harvard Medical School, Belmont, MA
* To whom correspondence should be addressed. E-mail: isacson{at}hms.harvard.edu.
Correspondence may also be addressed to Eva Hedlund at eva.hedlund@licr.ki.se
 |
Abstract |
|---|
Both fetal ventral mesencephalic (VM) and embryonic stem (ES) cell-derived dopamine neurons have been used successfully to correct behavioral responses in animal models of Parkinson's disease. However, grafts derived from fetal VM cells or from ES cells contain multiple cell types and the majority of these cells are not dopamine neurons. Isolation of ES cell-derived dopamine neurons and subsequent transplantation would elucidate both the capacity of these neurons to provide functional input and also further explore an efficient and safer use of ES cells for the treatment of Parkinson's disease. Towards this goal, we used a Pitx3-eGFP knock-in mouse ES (mES) cell line and fluorescence activated cell sorting (FACS) to select and purify midbrain dopamine neurons.
Initially, the dopaminergic marker profile of intact Pitx3-eGFP mES cultures was evaluated after differentiation in vitro. eGFP expression overlapped closely with Pitx3, Nurr1, Engrailed-1, Lmx1a, tyrosine hydroxylase (TH), L-aromatic amino acid decarboxylase (AADC) and vesicular monoamine transporter 2 (VMAT2) demonstrating that these cells were of a midbrain dopamine neuron character. Furthermore, postmitotic Pitx3-eGFP+ dopamine neurons, which constituted 2-5% of all live cells in the culture after dissociation, could be highly enriched to >90% purity by FACS and that these isolated neurons were viable, extended neurites and maintained a dopaminergic profile in vitro. Transplantation to 6-hydroxydopamine lesioned rats showed that an enriched dopaminergic population could survive and restore both amphetamine- and apomorphine-induced functions and the grafts contained large numbers of midbrain dopamine neurons, which innervated the host striatum.
______________________________________________________________________________
Author contributions: E.H.: Conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing; J.P.: collection and assembly of data, data analysis and interpretation; T.L.: collection and assembly of data, data analysis and interpretation; W.L.: collection and assembly of data, data analysis and interpretation; A.V.: collection and assembly of data, data analysis and interpretation; K.K.: Conception and design, financial support, data analysis and interpretation; O.I.: Conception and design, financial support, data analysis and interpretation, manuscript writing.
Key Words. Pitx3, transplantation, Parkinson's disease, stage-specific embryonic antigen-1 (SSEA-1), mouse embryonic stem cells
This article has been cited by other articles:
|
|
 |
|
  R. Sanchez-Pernaute, H. Lee, M. Patterson, C. Reske-Nielsen, T. Yoshizaki, K. C. Sonntag, L. Studer, and O. Isacson Parthenogenetic dopamine neurons from primate embryonic stem cells restore function in experimental Parkinson's disease Brain, August 1, 2008; 131(8): 2127 - 2139. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
| STEM CELLS | THE ONCOLOGIST | CME | ALPHAMED PRESS JOURNALS |
