title: Dopaminergic neurons derived from BG01V2, a variant of human embryonic stem cell line BG01.
abstract: OBJECTIVE
  Human embryonic stem cells (hESC) are considered a renewable source of dopamine producing neurons, and are of particular interest for their potential clinical use in Parkinson's disease. In this study, we characterized human dopaminergic neurons generated by stromal-derived inducing activity (SDIA) from BG01V2, a strain of human embryonic stem cell line, BG01, characterized by a chromosome 17 trisomy. Similar chromosomal changes have been repeatedly observed in hESC cultures in different laboratories, indicating the importance of chromosome 17 for growth and adaptation of hESC to culture.
  
  
  METHODS
  We investigated in vitro proliferation of differentiating cells using a BrDU incorporation assay, and monitored the cell population in long term cultures. Despite the cytogenetic abnormality, TH+ neurons were postmitotic at all stages of differentiation. After 30 days of differentiation, cell division ceased in 91% of the overall population of cells in the culture, indicating intact cell cycle regulation.
  
  
  RESULTS
  Expression of midbrain specific marker genes (Otx2, Pax5, Msx-1) showed differentiation of hESC-derived neural progenitor cells into midbrain specific dopamine neurons. These neurons expressed the dopamine transporter (DAT), and displayed functional DAT activity and electrical excitability.
  
  
  CONCLUSIONS
  TH+ cells derived from the BG01V2 hESC line using SDIA are postmitotic and have functional characteristics of normal dopaminergic neurons.
authors.affiliation: Development and Plasticity Section, Cellular Neurobiology Research Branch, Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, Department of Health and Human Services, Baltimore, MD, USA.
authors.forename: Tandis
authors.initials: T
authors.lastname: Vazin
authors.forename: Jia
authors.initials: J
authors.lastname: Chen
authors.forename: Charles E
authors.initials: CE
authors.lastname: Spivak
authors.forename: Rose
authors.initials: R
authors.lastname: Amable
authors.forename: Emily
authors.initials: E
authors.lastname: Gabitzsch
authors.forename: Chun-Ting
authors.initials: CT
authors.lastname: Lee
authors.forename: Carl R
authors.initials: CR
authors.lastname: Lupica
authors.forename: William J
authors.initials: WJ
authors.lastname: Freed
mesh_terms: D000199:Actins / Q000378:metabolism; D001973:Bromodeoxyuridine / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D049109:Cell Proliferation; D002886:Chromosomes, Human, Pair 17; D003042:Cocaine / Q000031:analogs & derivatives / Q000493:pharmacokinetics; D018920:Coculture Techniques; D004298:Dopamine / Q000378:metabolism*; D050483:Dopamine Plasma Membrane Transport Proteins / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000502:physiology*; D006801:Humans; D008564:Membrane Potentials / Q000502:physiology; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009474:Neurons / Q000502:physiology*; D051857:Otx Transcription Factors / Q000378:metabolism; D018408:Patch-Clamp Techniques / Q000379:methods; D011485:Protein Binding / Q000187:drug effects; D013997:Time Factors; D014316:Tritium / Q000493:pharmacokinetics; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
chemical_list: D000199:Actins; D050483:Dopamine Plasma Membrane Transport Proteins; C489788:MAP2 protein, human; D008869:Microtubule-Associated Proteins; D009419:Nerve Tissue Proteins; C497659:OTX2 protein, human; D051857:Otx Transcription Factors; D014316:Tritium; C021279:(1R-(exo,exo))-3-(4-fluorophenyl)-8-methyl-8- azabicyclo(3.2.1)octane-2-carboxylic acid, methyl ester; D014446:Tyrosine 3-Monooxygenase; D001973:Bromodeoxyuridine; D003042:Cocaine; D004298:Dopamine