Neural differentiation of mouse embryonic stem cells in serum-free monolayer culture

Wikrom Wongpaiboonwattana, Marios P. Stavridis (Lead / Corresponding author)

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    8 Citations (Scopus)
    124 Downloads (Pure)

    Abstract

    The ability to differentiate mouse embryonic stem cells (ESC) to neural progenitors allows the study of the mechanisms controlling neural specification as well as the generation of mature neural cell types for further study. In this protocol we describe a method for the differentiation of ESC to neural progenitors using serum-free, monolayer culture. The method is scalable, efficient and results in production of ~70% neural progenitor cells within 4-6 days. It can be applied to ESC from various strains grown under a variety of conditions. Neural progenitors can be allowed to differentiate further into functional neurons and glia or analyzed by microscopy, flow cytometry or molecular techniques. The differentiation process is amenable to time-lapse microscopy and can be combined with the use of reporter lines to monitor the neural specification process. We provide detailed instructions on media preparation and cell density optimization to allow the process to be applied to most ESC lines and a variety of cell culture vessels.
    Original languageEnglish
    Article number e52823
    JournalJoVE: Journal of Visualized Experiments
    Issue number99
    DOIs
    Publication statusPublished - 14 May 2015

    Fingerprint

    Embryonic Stem Cells
    Stem cells
    Cell culture
    Monolayers
    Microscopy
    Serum
    Microscopic examination
    Specifications
    Neuroglia
    Flow cytometry
    Flow Cytometry
    Stem Cells
    Cell Culture Techniques
    Cell Count
    Neurons
    Cell Line
    Mouse Embryonic Stem Cells

    Keywords

    • Embryonic stem cells
    • differentiation
    • screening
    • cell culture
    • N2B27
    • in vitro
    • monolayer
    • neural specification
    • developmental biology

    Cite this

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    title = "Neural differentiation of mouse embryonic stem cells in serum-free monolayer culture",
    abstract = "The ability to differentiate mouse embryonic stem cells (ESC) to neural progenitors allows the study of the mechanisms controlling neural specification as well as the generation of mature neural cell types for further study. In this protocol we describe a method for the differentiation of ESC to neural progenitors using serum-free, monolayer culture. The method is scalable, efficient and results in production of ~70{\%} neural progenitor cells within 4-6 days. It can be applied to ESC from various strains grown under a variety of conditions. Neural progenitors can be allowed to differentiate further into functional neurons and glia or analyzed by microscopy, flow cytometry or molecular techniques. The differentiation process is amenable to time-lapse microscopy and can be combined with the use of reporter lines to monitor the neural specification process. We provide detailed instructions on media preparation and cell density optimization to allow the process to be applied to most ESC lines and a variety of cell culture vessels.",
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    author = "Wikrom Wongpaiboonwattana and Stavridis, {Marios P.}",
    note = "Work was funded by a Development and Promotion of Science and Technology scholarship from the Thai Ministry for Education to W.W. and grants from Tenovus and the Anonymous Trust to M.P.S.",
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    T1 - Neural differentiation of mouse embryonic stem cells in serum-free monolayer culture

    AU - Wongpaiboonwattana, Wikrom

    AU - Stavridis, Marios P.

    N1 - Work was funded by a Development and Promotion of Science and Technology scholarship from the Thai Ministry for Education to W.W. and grants from Tenovus and the Anonymous Trust to M.P.S.

    PY - 2015/5/14

    Y1 - 2015/5/14

    N2 - The ability to differentiate mouse embryonic stem cells (ESC) to neural progenitors allows the study of the mechanisms controlling neural specification as well as the generation of mature neural cell types for further study. In this protocol we describe a method for the differentiation of ESC to neural progenitors using serum-free, monolayer culture. The method is scalable, efficient and results in production of ~70% neural progenitor cells within 4-6 days. It can be applied to ESC from various strains grown under a variety of conditions. Neural progenitors can be allowed to differentiate further into functional neurons and glia or analyzed by microscopy, flow cytometry or molecular techniques. The differentiation process is amenable to time-lapse microscopy and can be combined with the use of reporter lines to monitor the neural specification process. We provide detailed instructions on media preparation and cell density optimization to allow the process to be applied to most ESC lines and a variety of cell culture vessels.

    AB - The ability to differentiate mouse embryonic stem cells (ESC) to neural progenitors allows the study of the mechanisms controlling neural specification as well as the generation of mature neural cell types for further study. In this protocol we describe a method for the differentiation of ESC to neural progenitors using serum-free, monolayer culture. The method is scalable, efficient and results in production of ~70% neural progenitor cells within 4-6 days. It can be applied to ESC from various strains grown under a variety of conditions. Neural progenitors can be allowed to differentiate further into functional neurons and glia or analyzed by microscopy, flow cytometry or molecular techniques. The differentiation process is amenable to time-lapse microscopy and can be combined with the use of reporter lines to monitor the neural specification process. We provide detailed instructions on media preparation and cell density optimization to allow the process to be applied to most ESC lines and a variety of cell culture vessels.

    KW - Embryonic stem cells

    KW - differentiation

    KW - screening

    KW - cell culture

    KW - N2B27

    KW - in vitro

    KW - monolayer

    KW - neural specification

    KW - developmental biology

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    DO - 10.3791/52823

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    C2 - 26066640

    JO - JoVE: Journal of Visualized Experiments

    JF - JoVE: Journal of Visualized Experiments

    SN - 1940-087X

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