title: A protocol for the differentiation of human embryonic stem cells into dopaminergic neurons using only chemically defined human additives: Studies in vitro and in vivo.
abstract: Our ability to use human embryonic stem (hES) cells in cell replacement therapy for Parkinson's disease depends on the discovery of ways to simply and reliably differentiate a dopaminergic (DA) phenotype in these cells. Although several protocols exist for the differentiation of DA traits in hES, they involve the prolonged use of complex media with undefined components, cell conditioned media and/or co-culture with various cells, usually of animal origin. In this study, several well-characterized (H9, BG01) and several new uncharacterized (HUES7, HUES8) hES cell lines were studied for their capacity to differentiate into DA neurons in culture using a novel rapid protocol which uses only chemically-defined human-derived media additives and substrata. Within 3 weeks, cells from all 4 cell lines progressed from the undifferentiated state to beta-tubulin III positive cells expressing DA markers in vitro. Moreover, transplantation of these cells into the striata of 6-hydroxydopamine-treated rats at the neuronal progenitor stage resulted in the appearance of differentiated DA traits in vivo 2-3 weeks later.
authors.affiliation: Farber Institute for Neurosciences, Department of Neurology, Thomas Jefferson University Medical College, Suite 462-Jefferson Hospital for Neuroscience, 900 Walnut Street, Philadelphia, PA 19107, USA. lorraine.iacovitti@jefferson.edu
authors.forename: Lorraine
authors.initials: L
authors.lastname: Iacovitti
authors.forename: Angela E
authors.initials: AE
authors.lastname: Donaldson
authors.forename: Cheryl E
authors.initials: CE
authors.lastname: Marshall
authors.forename: Sokreine
authors.initials: S
authors.lastname: Suon
authors.forename: Ming
authors.initials: M
authors.lastname: Yang
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D003994:Bucladesine / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D002460:Cell Line; D002478:Cells, Cultured; D024141:Collagen Type IV / Q000494:pharmacology; D003342:Corpus Striatum / Q000503:physiopathology / Q000601:surgery; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology; D004298:Dopamine / Q000378:metabolism*; D004299:Dopamine beta-Hydroxylase / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D006085:Graft Survival / Q000502:physiology; D006801:Humans; D007381:Intermediate Filament Proteins / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism; D064231:Nestin; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D016627:Oxidopamine; D020734:Parkinsonian Disorders / Q000503:physiopathology / Q000628:therapy; D010455:Peptides / Q000494:pharmacology; D033581:Stem Cell Transplantation / Q000379:methods; D014404:Tubulin / Q000378:metabolism
chemical_list: D015415:Biomarkers; D024141:Collagen Type IV; D003470:Culture Media; D007381:Intermediate Filament Proteins; C577484:NES protein, human; D009419:Nerve Tissue Proteins; C577486:Nes protein, rat; D064231:Nestin; D010455:Peptides; D014404:Tubulin; D016222:Fibroblast Growth Factor 2; C008973:polyornithine; D003994:Bucladesine; D016627:Oxidopamine; D004299:Dopamine beta-Hydroxylase; D004298:Dopamine