Directed Differentiation of Dopaminergic Neuronal Subtypes from Human Embryonic Stem Cells
Open Access
- 1 June 2005
- journal article
- research article
- Published by Oxford University Press (OUP) in The International Journal of Cell Cloning
- Vol. 23 (6), 781-790
- https://doi.org/10.1634/stemcells.2004-0365
Abstract
How dopamine (DA) neuronal subtypes are specified remains unknown. In this study we show a robust generation of functional DA neurons from human embryonic stem cells (hESCs) through a specific sequence of application of fibroblast growth factor 8 (FGF8) and sonic hedgehog (SHH). Treatment of hESC‐derived Sox1+neuroepithelial cells with FGF8 and SHH resulted in production of tyrosine hydroxylase (TH)–positive neurons that were mostly bipolar cells, coexpression with γ‐aminobutyric acid, and lack of midbrain marker engrailed 1 (En1) expression. However, FGF8 treatment of precursor cells before Sox1 expression led to the generation of a similar proportion of TH+ neurons characteristic of midbrain projection DA neurons with large cell bodies and complex processes and coexpression of En1. This suggests that one mechanism of generating neuronal subtypes is temporal availability of morphogens to a specific group of precursors. The in vitro–generated DA neurons were electrophysiologically active and released DA in an activity‐dependent manner. They may thus provide a renewable source of functional human DA neurons for drug screening and development of sustainable therapeutics for disorders affecting the DA system.Keywords
This publication has 50 references indexed in Scilit:
- Specification of motoneurons from human embryonic stem cellsNature Biotechnology, 2005
- Generation of Tyrosine Hydroxylase Positive Neurons from Human Embryonic Stem Cells after Coculture with Cellular Substrates and Exposure to GDNFThe International Journal of Cell Cloning, 2004
- Engrailed genes are cell-autonomously required to prevent apoptosis in mesencephalic dopaminergic neuronsDevelopment, 2004
- Optimal conditions for in vivo induction of dopaminergic neurons from embryonic stem cells through stromal cell‐derived inducing activityJournal of Neuroscience Research, 2002
- Specification of catecholaminergic and serotonergic neuronsNature Reviews Neuroscience, 2002
- In vitro differentiation of transplantable neural precursors from human embryonic stem cellsNature Biotechnology, 2001
- Neural plate patterning: Upstream and downstream of the isthmic organizerNature Reviews Neuroscience, 2001
- Neuronal specification in the spinal cord: inductive signals and transcriptional codesNature Reviews Genetics, 2000
- Efficient generation of midbrain and hindbrain neurons from mouse embryonic stem cellsNature Biotechnology, 2000
- Embryonic Stem Cell Lines Derived from Human BlastocystsScience, 1998