Isolation and transplantation of dopaminergic neurons generated from mouse embryonic stem cells.
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.
Yoshizaki
Takahito
T
Department of Physiuology, Keio University School of Medicine, Tokyo, Japan.
Inaji
Motoki
M
Kouike
Hiroko
H
Shimazaki
Takuya
T
Sawamoto
Kazunobu
K
Ando
Kiyoshi
K
Date
Isao
I
Kobayashi
Kazuto
K
Suhara
Tetsuya
T
Uchiyama
Yasuo
Y
Okano
Hideyuki
H
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
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D014446:Tyrosine 3-Monooxygenase; D004298:Dopamine