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    Stem Cells. 2008 Jul;26(7):1705-12. Epub 2008 Apr 17.

    Lineage selection of functional and cryopreservable human embryonic stem cell-derived neurons.

    Ladewig J, Koch P, Endl E, Meiners B, Opitz T, Couillard-Despres S, Aigner L, Brüstle O.

    Source

    Institute of Reconstructive Neurobiology, Life & Brain Center, University of Bonn, Sigmund-Freud-Strasse 25, D-53105 Bonn, Germany.

    Abstract

    A major prerequisite for the biomedical application of human embryonic stem cells (hESC) is the derivation of defined and homogeneous somatic cell types. Here we present a human doublecortin (DCX) promoter-based lineage-selection strategy for the generation of purified hESC-derived immature neurons. After transfection of hESC-derived neural precursors with a DCX-enhanced green fluorescent protein construct, fluorescence-activated cell sorting enables the enrichment of immature human neurons at purities of up to 95%. Selected neurons undergo functional maturation and are able to establish synaptic connections. Considering that the applicability of purified hESC-derived neurons would largely benefit from an efficient cryopreservation technique, we set out to devise defined freezing conditions involving caspase inhibition, which yield post-thaw recovery rates of up to 83%. Combined with our lineage-selection procedure this cryopreservation technique enables the generation of human neurons in a ready-to-use format for a large variety of biomedical applications.

    PMID:
    18420830
    [PubMed - indexed for MEDLINE]
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