Embryonic Stem Cell Protocols

Volume 330 of the series Methods in Molecular Biology™ pp 33-54

Derivation and Characterization of Neural Cells From Embryonic Stem Cells Using Nestin Enhancer

  • Nibedita LenkaAffiliated withNational Center for Cell Science, Ganeshkhind

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The embryonic stem (ES) cells derived from the inner cell mass of the blastula stage embryo bear the complete repertoire of the complex organizational blueprint of an organism. These fascinating cells are bestowed with pluripotent characteristics and can be directed to differentiate into various lineages in vitro. Hence, these cells are being explored as an ideal in vitro model in gaining insight into early developmental events. Using the ES cell system, we have tried to investigate the early neurogenic proceedings. We have taken advantage of nestin enhancer-mediated cell trapping using the live reporter-based system. We monitored live the ES cell differentiation into neural lineage by following the enhanced green fluorescent protein expression profile in a number of stable ES cell clones generated by us in which the enhanced green fluorescent protein expression was regulated by the nestin enhancer. This strategy has helped us in both qualitative and quantitative detection and characterization of neural progenitor population and the differentiated progenies.

Key Words

ES cells glia live reporter nestin neural stem cells neurogenesis neuron