Astrocytic responses to DNA delivery using Nucleofection

Hakan Muyderman, Wai Yew, Benjaporn Homkajorn, Neil Sims

    Research output: Contribution to journalArticlepeer-review

    3 Citations (Scopus)

    Abstract

    Nucleofection is a powerful non-viral transfection technique that can deliver plasmid DNA with high efficiency to cells that are traditionally difficult to transfect. In this study, we demonstrate that nucleofection of astrocytes grown in primary cell culture resulted in 76 ± 9% transfected cells and low cytotoxicity. However, the nucleofected astrocytes showed a reduced re-attachment to the growth media when replated and subsequent impairment of proliferation. This led to substantially decreased cell densities during the initial 72 h following transfection. Furthermore, these cells were less efficient at producing wound closure in a scratch model of injury. Nucleofection also resulted in the generation of a small proportion of polynucleated cells. The findings demonstrate that nucleofection provides a valuable technique for delivering DNA to astrocytes in culture. However, considerable care is needed in designing and interpreting such studies because of long-lasting changes induced in key properties of these cells by the nucleofection process.

    Original languageEnglish
    Pages (from-to)1771-1779
    Number of pages9
    JournalNeurochemical Research
    Volume35
    Issue number11
    DOIs
    Publication statusPublished - Nov 2010

    Keywords

    • Astrocytes
    • Nucleofection
    • Scratch wound injury
    • Transfection

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