title: Isolation and transplantation of dopaminergic neurons generated from mouse embryonic stem cells.
abstract: 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.
authors.affiliation: Department of Physiuology, Keio University School of Medicine, Tokyo, Japan.
authors.forename: Takahito
authors.initials: T
authors.lastname: Yoshizaki
authors.forename: Motoki
authors.initials: M
authors.lastname: Inaji
authors.forename: Hiroko
authors.initials: H
authors.lastname: Kouike
authors.forename: Takuya
authors.initials: T
authors.lastname: Shimazaki
authors.forename: Kazunobu
authors.initials: K
authors.lastname: Sawamoto
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Ando
authors.forename: Isao
authors.initials: I
authors.lastname: Date
authors.forename: Kazuto
authors.initials: K
authors.lastname: Kobayashi
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Suhara
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Uchiyama
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Okano
mesh_terms: D000818:Animals; D001522:Behavior, Animal; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods; D003342:Corpus Striatum / Q000378:metabolism / Q000637:transplantation; D004195:Disease Models, Animal; D004298:Dopamine / Q000378:metabolism*; D004622:Embryo, Mammalian; D005434:Flow Cytometry / Q000379:methods; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000378:metabolism; D051379:Mice; D009474:Neurons / Q000378:metabolism / Q000637:transplantation*; D010300:Parkinson Disease / Q000601:surgery; D020127:Recovery of Function; D033581:Stem Cell Transplantation; D017154:Stromal Cells / Q000502:physiology; D013997:Time Factors; D014162:Transfection / Q000379:methods; D019737:Transplants; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
chemical_list: D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D014446:Tyrosine 3-Monooxygenase; D004298:Dopamine