Isolation and transplantation of dopaminergic neurons generated from mouse embryonic stem cells

Neurosci Lett. 2004 Jun 3;363(1):33-7. doi: 10.1016/j.neulet.2004.03.074.


Embryonic stem (ES) cells differentiate into dopamine (DA)-producing neurons when co-cultured with PA6 stromal cells, but the resulting cultures contain a variety of unidentified cells. In order to label live DA neurons in mixed populations, we introduced a GFP reporter under the control of the tyrosine hydroxylase (TH) gene promoter into ES cells. GFP expression was observed in TH-immunoreactive cells that differentiated from the ES cells that carried the TH-GFP reporter gene. DA neurons expressing GFP were sorted from the mixed cell population by fluorescence-activated cell sorting of cells exhibiting GFP fluorescence, and the sorted GFP(+) cells obtained were transplanted into a rat model of Parkinson's disease. Some of these cells survived and innervated the host striatum, resulting in a partial recovery from parkinsonian behavioral defects. This strategy of isolation and transplantation of ES-cell-derived DA neurons should be useful for cellular and molecular studies of DA neurons and for clinical application in the treatment of Parkinson's disease.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Behavior, Animal
  • Cell Separation / methods
  • Cells, Cultured
  • Coculture Techniques / methods
  • Corpus Striatum / metabolism
  • Corpus Striatum / transplantation
  • Disease Models, Animal
  • Dopamine / metabolism*
  • Embryo, Mammalian
  • Flow Cytometry / methods
  • Green Fluorescent Proteins
  • Luminescent Proteins / metabolism
  • Mice
  • Neurons / metabolism
  • Neurons / transplantation*
  • Parkinson Disease / surgery
  • Recovery of Function
  • Stem Cell Transplantation*
  • Stromal Cells / physiology
  • Time Factors
  • Transfection / methods
  • Transplants*
  • Tyrosine 3-Monooxygenase / metabolism


  • Luminescent Proteins
  • Green Fluorescent Proteins
  • Tyrosine 3-Monooxygenase
  • Dopamine