A method for rapid derivation and propagation of neural progenitors from human embryonic stem cells.

• Neuronal loss is a common feature of many neurological disorders, including stroke, Parkinson's disease, Alzheimer's disease and traumatic brain injury. Human embryonic stem cell (hESC)-derived neural progenitors (NPs) may provide new ways of treatment for several diseases and injuries in the brain, as well as enhance our understanding of early human development. Here we report a method for rapid generation of proliferating NPs from feeder free cultures of undifferentiated hESC. In this rapid and simple protocol, NPs are derived by seeding undifferentiated hESC on adherent surfaces of laminin or gelatine with normal hESC culturing medium and with the addition of basic fibroblast growth factor. After the first passage, adherent monolayer progenitors are derived that express early neuroectodermal and progenitor markers, such as Nestin, Sox1, Sox2, Sox3, Internexin, Musashi-1, NCAM, and Pax6. This novel protocol renders hESC suitable for large scale progenitor production and long-term propagation, and the progenitors have the capacity to differentiate in vitro into all three neural lineages (neurons, astrocytes and oligodendrocytes). This method allows rapid, cost-efficient production of expandable progenitors that may be a source of cells for the restoration of cellular and functional loss after neurodegeneration and/or provide a useful source of progenitor cells for studying early brain development.

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