Genetic modification of dendritic cells through the directed differentiation of embryonic stem cells

Paul J. Fairchild, Kathleen F. Nolan, Herman Waldmann

    Research output: Contribution to journalArticlepeer-review

    Abstract

    Recent years have witnessed a progressive acceptance of the dual role played by dendritic cells (DC) in the initiation of immune responses and their specific attenuation through the induction of immunological tolerance. Nevertheless, as terminally differentiated cells of the myeloid lineage, DC share with macrophages an inherent resistance to genetic modification, greatly restricting strategies available for studying their physiology and function. Consequently, little is known of the molecular interactions provided by DC that underlie the critical decision between tolerance and immunity. Embryonic stem (ES) cells are, by contrast, relatively amenable to genetic modification. Furthermore, their propensity for self-renewal, one of the cardinal features of a stem cell, permits cloning at the single cell level and the rational design of ES cell lines, uniformly expressing a desired, mutant phenotype. Here, we describe how another defining property of ES cells, their demonstrable pluripotency, may be harnessed for their directed differentiation along the DC pathway, enabling the generation of limitless numbers of DC faithfully expressing candidate genes of interest. The protocols we outline in this chapter may, therefore, offer new opportunities for dissecting the biology of DC and the molecular basis of their unique properties. © Humana Press Inc.
    Original languageEnglish
    Pages (from-to)59-72
    Number of pages13
    JournalMethods in Molecular Biology
    Volume380
    DOIs
    Publication statusPublished - 25 Apr 2007

    Keywords

    • Dendritic cell
    • Directed differentiation
    • Embryonic stem cell
    • Genetic modification
    • Tolerance

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