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      Conversion of embryonic stem cells into neuroectodermal precursors in adherent monoculture.

      Nature biotechnology
      Animals, Cell Aggregation, drug effects, physiology, Cell Differentiation, Cell Line, Culture Media, pharmacology, Ectoderm, metabolism, Growth Inhibitors, Interleukin-6, Leukemia Inhibitory Factor, Lymphokines, Mice, Neuroglia, cytology, Neurons, Pluripotent Stem Cells, Stem Cells

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          Abstract

          Mouse embryonic stem (ES) cells are competent for production of all fetal and adult cell types. However, the utility of ES cells as a developmental model or as a source of defined cell populations for pharmaceutical screening or transplantation is compromised because their differentiation in vitro is poorly controlled. Specification of primary lineages is not understood and consequently differentiation protocols are empirical, yielding variable and heterogeneous outcomes. Here we report that neither multicellular aggregation nor coculture is necessary for ES cells to commit efficiently to a neural fate. In adherent monoculture, elimination of inductive signals for alternative fates is sufficient for ES cells to develop into neural precursors. This process is not a simple default pathway, however, but requires autocrine fibroblast growth factor (FGF). Using flow cytometry quantitation and recording of individual colonies, we establish that the bulk of ES cells undergo neural conversion. The neural precursors can be purified to homogeneity by fluorescence activated cell sorting (FACS) or drug selection. This system provides a platform for defining the molecular machinery of neural commitment and optimizing the efficiency of neuronal and glial cell production from pluripotent mammalian stem cells.

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