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Various Nucleofector™ Kits and corresponding Amaxa™ Optimized Protocols are available for the transfection of rat NSCs using the different Nucleofection™ Platforms. For the transfection of rat 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. Rat NSC specific kits are available for the Nucleofector™ II/2b Device.
Nucleofection™ of rat NSCs
Primary NSC isolated from rat embryos (E14) were transfected by Nucleofection™ using a plasmid encoding enhanced green fluorescent protein eGFP under control of an EF1alpha promoter (pcDNAEF1-eGFP). Post Nucleofection™ cells were cultured with bFGF for 2 days, then for 5 additional days without bFGF to differentiate into neurons and astrocytes. Cells were analyzed 2 days (left) and 7 days (right) post Nucleofection™ by fluorescence microscopy.
(Courtesy of S.H. Lee, College of Medicine, Dept. of Biochemistry, Hanyang University, Seoul, South Korea)