Increasing cellular lifespan with a flow system in organotypic culture of the Laterodorsal Tegmentum (LDT)

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    Abstract

    Organotypic brain culture is an experimental tool widely used in neuroscience studies. One major
    drawback of this technique is reduced neuronal survival across time, which is likely exacerbated by the
    loss of blood flow. We have designed a novel, tube flow system, which is easily incorporated into the
    commonly-used, standard semi-permeable membrane culture methodology which has significantly
    enhanced neuronal survival in a brain stem nucleus involved in control of motivated and arousal states:
    the laterodorsal tegmental nucleus (LDT). Our automated system provides nutrients and removes
    waste in a comparatively aseptic environment, while preserving temperature, and oxygen levels. Using
    immunohistochemistry and electrophysiology, our system was found superior to standard techniques
    in preserving tissue quality and survival of LDT cells for up to 2 weeks. In summary, we provide
    evidence for the first time that the LDT can be preserved in organotypic slice culture, and further, our
    technical improvements of adding a flow system, which likely enhanced perfusion to the slice, were
    associated with enhanced neuronal survival. Our perfusion system is expected to facilitate organotypic
    experiments focused on chronic stimulations and multielectrode recordings in the LDT, as well as
    enhance neuronal survival in slice cultures originating from other brain regions.
    Original languageEnglish
    JournalScientific Reports
    Pages (from-to)1 - 11
    Number of pages11
    ISSN2045-2322
    Publication statusPublished - 1 Dec 2019

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