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

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

Nature biotechnology (2003-01-14)
Qi-Long Ying, Marios Stavridis, Dean Griffiths, Meng Li, Austin Smith
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
NDiff Neuro-27 Medium Supplement (100x), NDiff Neuro-27 Medium Supplement is a serum-free supplement containing antioxidants & other factors that has been developed for the in vitro propagation & maintenance of undifferentiated murine embryonic stem (ES) cells in serum free medium.
Pricing and availability is not currently available.
Sigma-Aldrich
NDiff Neuro-2 Medium Supplement (200x), NDiff Neuro-2 Medium Supplement is a serum-free, N2-like supplement for the in vitro differentiation of murine embryonic stem (ES) cells into post-mitotic neurons particularly via monolayer differentiation.
Pricing and availability is not currently available.