Conversion of embryonic stem cells into neuroectodermal precursors in adherent monoculture

Qi-Long Ying, Marios Stavridis, Dean Griffiths, Meng Li, Austin Smith

    Research output: Contribution to journalArticle

    1030 Citations (Scopus)

    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.
    Original languageEnglish
    Pages (from-to)183-6
    Number of pages4
    JournalNature Biotechnology
    Volume21
    Issue number2
    DOIs
    Publication statusPublished - 2003

    Fingerprint

    Embryonic Stem Cells
    Stem cells
    Flow Cytometry
    Pluripotent Stem Cells
    Fibroblast Growth Factors
    Coculture Techniques
    Cells
    Neuroglia
    Pharmaceutical Preparations
    Flow cytometry
    Transplantation
    Fibroblasts
    Sorting
    Drug products
    Machinery
    Screening
    Agglomeration
    Fluorescence
    Population
    Specifications

    Cite this

    Ying, Qi-Long ; Stavridis, Marios ; Griffiths, Dean ; Li, Meng ; Smith, Austin. / Conversion of embryonic stem cells into neuroectodermal precursors in adherent monoculture. In: Nature Biotechnology. 2003 ; Vol. 21, No. 2. pp. 183-6.
    @article{8bc681eb41b8475dba765688126b5393,
    title = "Conversion of embryonic stem cells into neuroectodermal precursors in adherent monoculture",
    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.",
    author = "Qi-Long Ying and Marios Stavridis and Dean Griffiths and Meng Li and Austin Smith",
    year = "2003",
    doi = "10.1038/nbt780",
    language = "English",
    volume = "21",
    pages = "183--6",
    journal = "Nature Biotechnology",
    issn = "1087-0156",
    publisher = "Nature Publishing Group",
    number = "2",

    }

    Conversion of embryonic stem cells into neuroectodermal precursors in adherent monoculture. / Ying, Qi-Long; Stavridis, Marios; Griffiths, Dean; Li, Meng; Smith, Austin.

    In: Nature Biotechnology, Vol. 21, No. 2, 2003, p. 183-6.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Conversion of embryonic stem cells into neuroectodermal precursors in adherent monoculture

    AU - Ying, Qi-Long

    AU - Stavridis, Marios

    AU - Griffiths, Dean

    AU - Li, Meng

    AU - Smith, Austin

    PY - 2003

    Y1 - 2003

    N2 - 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.

    AB - 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.

    U2 - 10.1038/nbt780

    DO - 10.1038/nbt780

    M3 - Article

    C2 - 12524553

    VL - 21

    SP - 183

    EP - 186

    JO - Nature Biotechnology

    JF - Nature Biotechnology

    SN - 1087-0156

    IS - 2

    ER -