Toxic Effects of Lipid-Mediated Gene Transfer in Ventral Mesencephalic Explant Cultures

Authors

  • Matthias Bauer,

    1. GSF – National Research Center for Environment and Health, Institute for Human Genetics, Ingolstädter Landstraße 1, D-85764 Neuherberg, Germany
    2. Department of Neurology, Technical University of Munich, Nöhlstr. 28, 81675 München, Germany
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  • Bjarne Winther Kristensen,

    1. Department of Anatomy and Neurobiology, University of Southern Denmark, Odense, Winsløwparken 21, DK-5000 Odense C, Denmark
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  • Morten Meyer,

    1. Department of Anatomy and Neurobiology, University of Southern Denmark, Odense, Winsløwparken 21, DK-5000 Odense C, Denmark
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  • Thomas Gasser,

    1. Department of Neurology, University of Tübingen, Hoppe-Seyler Str. 3, D-72076 Tübingen, Germany
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  • Hans Ruedi Widmer,

    1. Department of Neurosurgery, Inselspital, University of Bern, Freiburgstraße, 3010 Bern, Switzerland
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  • Jens Zimmer,

    1. Department of Anatomy and Neurobiology, University of Southern Denmark, Odense, Winsløwparken 21, DK-5000 Odense C, Denmark
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  • Marius Ueffing

    Corresponding author
    1. GSF – National Research Center for Environment and Health, Institute for Human Genetics, Ingolstädter Landstraße 1, D-85764 Neuherberg, Germany
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Author for correspondence: Marius Ueffing, GSF – National Research Center for Environment and Health, Institute for Human Genetics, Ingolstädter Landstraße 1, D-85764 Neuherberg, Germany (fax +4989 3187 3297, e-mail marius.ueffing@gsf.de).

Abstract

Abstract: Adverse effects of cDNA and oligonucleotide delivery methods have not yet been systematically analyzed. We introduce a protocol to monitor toxic effects of two non-viral lipid-based gene delivery protocols using CNS primary tissue. Cell membrane damage was monitored by quantifying cellular uptake of propidium iodide and release of cytosolic lactate dehydrogenase to the culture medium. Using a liposomal transfection reagent, cell membrane damage was already seen 24 hr after transfection. Nestin-positive target cells, which were used as morphological correlate, were severely diminished in some areas of the cultures after liposomal transfection. In contrast, the non-liposomal transfection reagent revealed no signs of toxicity. This approach provides easily accessible information of transfection-associated toxicity and appears suitable for prescreening of transfection reagents.

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