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Original Article |
1 McLean Hospital/Harvard University Udall Parkinson's Disease Research Center of Excellence; Neuroregeneration Laboratories, McLean Hospital, 115 Mill St., Belmont, MA 02478
2 Laboratory of Stem Cell and Tumor Biology, Division of Neurosurgery, Memorial Sloan-Kettering Cancer Center, 1275 York Ave, Box 256, New York, NY 10021
3 Neuroregeneration Laboratories, McLean Hospital, 115 Mill St., Belmont, MA 02478; Stem Cell Research Institute, DIBIT, S. Raffaele Hospital, Via Olgettina 58, 20132 Milano
* To whom correspondence should be addressed. E-mail: rosario_pernaute{at}hms.harvard.edu.
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Abstract |
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Dopamine neurons can be derived from human and primate embryonic stem (ES) cells in vitro. An ES cell-based replacement therapy for patients with Parkinson's disease requires that in vitro generated neurons maintain their phenotype in vivo. Other critical issues relate to their proliferative capacity and risk of tumor formation, and the capability of migration and integration in the adult mammalian brain. Neural induction was achieved by co-culture of primate parthenogenetic ES cells (Cyno-1) with stromal cells followed by sequential exposure to midbrain patterning and differentiation factors to favor dopamine phenotypic specification. Differentiated ES cells were treated with mitomycin C and transplanted into adult immunosuppressed rodents and into a primate (allograft) without immunosuppression. A small percentage of dopamine neurons survived in both rodent and primate hosts for the entire term of the study (4 and 7 months, respectively). Other neuronal and glial populations derived from Cyno1 ES cells showed, in vivo, phenotypic characteristics, growth and migration patterns similar to fetal primate transplants and a majority of cells (>80%) expressed the forebrain transcription factor brain factor 1. No teratoma formation was observed. In this study we demonstrate long-term survival of dopamine neurons obtained in vitro from primate ES cells. Optimization of differentiation, cell selection and cell transfer is required for functional studies of ES-derived dopamine neurons for future therapeutic applications.
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