INDUCTION OF DOPAMINE NEURONS FROM EMBRYONIC STEM CELLS

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. To explore the possibility of generating dopamine neurons from an unlimited source of human and nonhuman primate embryonic stem (ES) cells to study potential cell replacement therapies for Parkinson's disease. We established a strategy that directs the pluripotent stem cells to a neural fate. This process entails aggregation and culture of ES cells in suspension, followed by differentiation of the ES cell aggregates in an adherent condition with a cocktail containing fibroblast growth factor 2 (FGF2) for 5-7 days. Neural precursor cells displayed a characteristic columnar morphology, organized into neural tube-like rosette formations, and expressed neuroepithelial marker, nestin and NCAM. These neural precursor cells were separated from surrounding non-neural cells to 95% purity by differential enzymatic response and differential adhesion. The isolated neural precursor cells differentiated into neurons and glial cells after withdraw of FGF2 in vitro. Ongoing study is to guide the ES-derived neural precursors further to dopamine neurons and examine the function of the dopamine neurons in a rat model of Parkinson's disease. This Research used WNPRC Stem Cell Resources and federally approved hES cell lines. FUNDING NOTE: Private funding.