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Stem Cells 2004;22:669-674 www.StemCells.com
© 2004 AlphaMed Press

RAPID COMMUNICATION

Generation of Tyrosine Hydroxylase Positive Neurons from Human Embryonic Stem Cells after Coculture with Cellular Substrates and Exposure to GDNF

Kimberley A. Buytaert-Hoefen, Enrique Alvarez, Curt R. Freed

Division of Clinical Pharmacology, Department of Medicine, University of Colorado Health Sciences Center, Denver, Colorado, USA

Key Words. Parkinson’s disease • Astrocytes • PA6 cells • Dopamine neurons

Correspondence: Curt R. Freed, M.D., University of Colorado Health Sciences Center, 4200 East Ninth Avenue, C237, Denver, Colorado 80262, USA. Telephone: 303-315-8455; Fax: 303-315-3272; e-mail: Curt.Freed{at}UCHSC.edu

Tyrosine hydroxylase (TH)–positive neurons were generated from human embryonic stem (hES) cells by coculturing on astrocytes or PA6 stromal cells. After 3 to 4 weeks in culture, TH-positive cells with neuronal morphology developed. Coculture with astrocytes from the embryonic striatum produced a larger number of TH-positive cells than did coculture with astrocytes from embryonic mesencephalon (329 ± 149 versus 33 ± 16 TH-positive cells per well, p < .05). In other experiments using PA6 cells as a substrate, glial-derived neurotrophic factor (GDNF) was added to the media of differentiating hES cells, and this led to a doubling of the number of TH-positive cells (PA6: 443 ± 105 TH-positive cells per well versus PA6 + GDNF: 934 ± 136, p < .05). We conclude that substrates of striatal astrocytes and PA6 cells can promote differentiation of human embryonic stem cells to a TH-positive phenotype and that GDNF can increase the number of cells expressing that phenotype.




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