Generation of tyrosine hydroxylase positive neurons from human embryonic stem cells after coculture with cellular substrates and exposure to GDNF.
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.
Buytaert-Hoefen
Kimberley A
KA
Department of Medicine, University of Colorado Health Sciences Center, Denver 80262, USA.
Alvarez
Enrique
E
Freed
Curt R
CR
D001253:Astrocytes / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D018920:Coculture Techniques; D003342:Corpus Striatum / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D017077:Culture Media, Conditioned / Q000378:metabolism / Q000494:pharmacology; D004298:Dopamine / Q000096:biosynthesis*; D051100:Glial Cell Line-Derived Neurotrophic Factor; D006801:Humans; D008636:Mesencephalon / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D009414:Nerve Growth Factors / Q000378:metabolism / Q000494:pharmacology*; D009474:Neurons / Q000187:drug effects / Q000201:enzymology*; D010300:Parkinson Disease / Q000628:therapy; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000187:drug effects / Q000201:enzymology*; D033581:Stem Cell Transplantation / Q000379:methods; D017154:Stromal Cells / Q000166:cytology; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism*
D017077:Culture Media, Conditioned; C495039:GDNF protein, human; D051100:Glial Cell Line-Derived Neurotrophic Factor; D009414:Nerve Growth Factors; D014446:Tyrosine 3-Monooxygenase; D004298:Dopamine