Approaches which differentiate embryonic stem (ES) cells into neurons have recently garnered greater attention mainly due to their importance in physiological research and possible applications in regenerative medicine. However, much effort is being marshaled to generate uniform neuronal populations, but a completely reliable method has yet to be developed. Herein, methods which have been reported are summarized and discussed. Based on the principle of isolating glutamatergic neurons, they can be classifi ed into approaches of immunoisolation, gene manipulation, and ectoderm dissociation. Each approach claims to be able to obtain uniform glutamatergic neurons. This chapter summarizes the three methods, describes detailed isolation procedures, and discusses functionally related studies. Some concerns with and disadvantages of each method still remain as protocols are further developed for clinical application. Nevertheless, establishing these methods has confi rmed that neurons derived from ES cell differentiation are excellent cellular models for investigating properties related to the central nervous system. They also have great potential for applications to regenerative medicine.
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