| Summary: | Human embryonic stem cells (hESCs) are a potential source of defined cell types for studying early human development and application in regenerative medicine. Realising this potential requires a number of challenges to be overcome. The experimental findings reported represent a systematic approach in establishing controlled and standardised conditions for differentiating hESCs down the neural lineage, and characterising neural derivatives both <i>in vitro</i> and <i>in vivo.</i> Human embryonic stem cell cultures were established from two independently-derived liens, H9 and UES9. A novel, efficient method for propagating hESCs is described, avoiding the use of enzymatic products which may lead to karyotypic instability. Controlled neuroectodermal differentiation is demonstrated using a chemically defined system over a period of 16 days, and this process is shown to be dependent on endogenous fibroblast growth factor (FGF) signalling. Neural progenitors generated with this system are subsequently expanded for over 180 days and shown to retain neural stem cell (NSC) identity at the clonal level. Evidence is provided that hESC-derived NSCs follow a developmentally predictable timecourse of neurogenesis followed by gliogenesis, and their <i>in vitro</i> and <i>in vivo</i> behaviour is characterised with respect to temporal maturation and phenotypic potential.
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