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Various Nucleofector™ Kits and corresponding Amaxa™ Optimized Protocols are available for the transfection of mouse NSCs using the different Nucleofection™ Platforms. For the transfection of mouse NSCs in the 4D-Nucleofector™ and 96-well Shuttle™ System we recommend using the Primary Cell Optimization Kits and the respective optimization protocols. Optimal Nucleofection™ Conditions are transferable between these two systems. Mouse NSC specific kits are available for the Nucleofector™ II/2b Device.
Nucleofection™ of mouse NSCs
Mouse neural stem cells were isolated from spinal cords of mouse embryos (E14-15) and cultured for 3 passages in medium containing EGF and FGF. Cells were transfected by Nucleofection™ a plasmid encoding the enhanced green fluorescent protein eGFP. After Nucleofection™ the cells were cultured in medium containing 1% FCS to initiate differentiation into neurons and astrocytes. 72 hours post Nucleofection™ cells were analyzed by fluorescence microscopy.
(Courtesy of I. Richard and U. Bartsch, Center for Molecular Neurobiology, Hamburg, Germany)