Generation of tyrosine hydroxylase positive neurons from human embryonic stem cells after coculture with cellular substrates and exposure to GDNF

Stem Cells. 2004;22(5):669-74. doi: 10.1634/stemcells.22-5-669.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Astrocytes / metabolism
  • Cell Differentiation / drug effects
  • Cell Differentiation / physiology*
  • Cell Proliferation / drug effects
  • Cell Shape / drug effects
  • Cell Shape / physiology
  • Cells, Cultured
  • Coculture Techniques
  • Corpus Striatum / cytology
  • Corpus Striatum / embryology
  • Corpus Striatum / metabolism
  • Culture Media, Conditioned / metabolism
  • Culture Media, Conditioned / pharmacology
  • Dopamine / biosynthesis*
  • Glial Cell Line-Derived Neurotrophic Factor
  • Humans
  • Mesencephalon / cytology
  • Mesencephalon / embryology
  • Mesencephalon / metabolism
  • Nerve Growth Factors / metabolism
  • Nerve Growth Factors / pharmacology*
  • Neurons / drug effects
  • Neurons / enzymology*
  • Parkinson Disease / therapy
  • Phenotype
  • Pluripotent Stem Cells / drug effects
  • Pluripotent Stem Cells / enzymology*
  • Stem Cell Transplantation / methods
  • Stromal Cells / cytology
  • Tyrosine 3-Monooxygenase / metabolism*

Substances

  • Culture Media, Conditioned
  • GDNF protein, human
  • Glial Cell Line-Derived Neurotrophic Factor
  • Nerve Growth Factors
  • Tyrosine 3-Monooxygenase
  • Dopamine