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W584:Schroeder:AdherentNucleofection

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Efficient High-Throughput Transfection of Neuronal Networks Using NucleofectionTM

Schroeder, Jenny; Lenz, Dietmar; Lorbach, Elke; Koblizek, Thomas; Heinze, Andreas; Toell, Andrea; Andretta, Gina; Buesch, Stefanie; Domzalski, Sandra; Litzenberger, Daniela; Schaepermeier, Sabine; Spicker, Sonja; Spottke, Nicole; Mueller-Hartmann, Herbert; Lonza Cologne GmbH, Nattermannallee 1, 50829 Cologne, GERMANY

Efficient delivery of biomolecules, such as DNA, into neural cells is a promising tool to study a broad spectrum of cellular functions. Primary mammalian neural cells derived from solid tissue can be cultured in vitro and exhibit pivotal functions when grown in adherence. For the transfection of adherent neurons a variety of non-viral methods exist which do not achieve high transfection efficiencies.
Here we present an advanced application of the NucleofectorTM Technology for transfecting attached networks of primary mammalian neurons. This has been achieved by the development of a modified version of the NucleocuvetteTM Modules for use with the 96-well ShuttleTM System or the new 4D-NucleofectorTM System. The wells of these modules are designed for cultivation, NucleofectionTM and in situ analysis of adherent neural cells. This allows continuous cultivation of the neuronal cells for up to several weeks by eliminating the need for enzymatic or mechanical removal of the cells from their growth substrate. Furthermore, the cells can be transfected and analyzed at various time points during culture and thus maturation stages in vitro.
With up to 70% transfection efficiency and high viability for various primary neuron types adherent transfection using the new NucleocuvetteTM AD Modules provides a new tool for studying primary mammalian neurons without interruption of their physiological state.
Additionally, a complementary system allowing the adherent NucleofectionTM of neuronal networks in conventional 24-well tissue-culture plates and on cover slips will be presented.


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