( ...)Efficient DNA transfection of primary CNS neurons using TransMessenger ,,, Transfection Reagent

Frank Narz, Silke Janhsen, and Ute Krger
QIAGEN GmbH, Hilden, Germany

Primary neurons are extremely difficult to transfect, making genetic modification of neuronal cells by delivery of either siRNA or plasmid DNA challenging (1). Recently, Krichevsky and Kosik reported silencing of endogenous genes in primary rat hippocampal and cortical neurons using QIAGEN siRNA and TransMessenger Transfection Reagent (2,3 ).

In this study, we investigated whether TransMessenger Reagent (originally developed for RNA transfection of eukaryotic cells) would provide efficient transfection of DNA. We generated cultures of primary cortical neurons using CryoCells from rat brain cortex (QBM Cell Science, www.qbmcellscience.com), which contain a mixture of neuronal and glial cells. The cultured cells expressed characteristic markers and showed typical morphology and differentiation. We transfected the cells with two plasmids, one encoding green fluorescent protein (GFP) and the other encoding -galactosidase (-gal). No obvious signs of toxicity were detected after transfection. The combination of TransMessenger Reagent and CryoCells allowed efficient transfection of DNA into primary cortical neurons.

Materials and methods

CryoCells, Rat Brain Cortex (QBM Cell Science) were cultured according to the manufacturer's instructions. Briefly, to start a culture, a vial of CryoCells (containing 4 x 106 viable cells before freezing) was thawed and the cells were plated in Neurobasal Medium with B27 Supplement
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