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

• Published in: Neuroscience letters 2004-06, Vol.363 (1), p.33-37
• Main Authors: Yoshizaki, Takahito, Inaji, Motoki, Kouike, Hiroko, Shimazaki, Takuya, Sawamoto, Kazunobu, Ando, Kiyoshi, Date, Isao, Kobayashi, Kazuto, Suhara, Tetsuya, Uchiyama, Yasuo, Okano, Hideyuki
• Format: Article
• Language: English
• Subjects: Animals; Behavior, Animal; Biological and medical sciences; Cell Separation - methods; Cells, Cultured; Coculture Techniques - methods; Corpus Striatum - metabolism; Corpus Striatum - transplantation; Disease Models, Animal; Dopamine - metabolism; Dopamine neuron; Embryo, Mammalian; Embryonic stem cells; Flow Cytometry - methods; Fundamental and applied biological sciences. Psychology; Green Fluorescent Proteins; Luminescent Proteins - metabolism; Mice; Neurons - metabolism; Neurons - transplantation; Parkinson Disease - surgery; Recovery of Function; Stem Cell Transplantation; Stromal Cells - physiology; Stromal-cell derived inducing activity (SDIA); TH-GFP; Time Factors; Transfection - methods; Transplantation; Transplants; Tyrosine 3-Monooxygenase - metabolism; Vertebrates: nervous system and sense organs
• ISSN: 0304-3940; 1872-7972
• 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-GFPreporter 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.