Abstract
Abstract Cell lines isolated from tumors are an important tool to study cancer in vitro. They can be used for drug development as well as for understanding the basic mechanisms underlying cancer. Transfection of cancer cell lines with different molecules like plasmid DNA, siRNA or mRNA is often an integral part for this kind of research. Lonza's 4D-Nucleofector™ System is a modular system for the efficient transfection of primary cells and cell lines with a variety of substrates including plasmid DNA and siRNA. The 4D-Nucleofector™ X Unit supports the Nucleofection™ in two different formats. The aluminum-free 20µl Nucleocuvette™ Strip allows the transfection of low cell numbers down to 2x104 cells per reaction. As 16 reactions can be performed in parallel it is well suited for optimizing Nucleofection™ Conditions for cells lacking a ready-to-use Optimized Protocol. For higher cell numbers of up to 2x107 cells per reaction the same Nucleofection™ Conditions can be applied in the 100μl single Nucleocuvette™ Vessel. For higher throughput needs the 96-well Shuttle™ Add-on can be connected to the 4D-Nucleofector™ System. With this add-on six 20µl Nucleocuvette™ Strips can be processed in parallel allowing for screening applications or accelerating the optimization of transfection parameters for many cell types. In this study we used the 4D-Nucleofector™ System in combination with the 96-well Shuttle™ Add-on for optimizing transfection conditions for multiple cancer cell lines. An exemplary optimization process is depicted for the human prostate carcinoma cell line DU 145 and for the human colorectal adenocarcinoma cell line COLO 205. Optimization steps included the selection of the appropriate Nucleofector™ Solution, Nucleofector™ Program and plasmid DNA concentration. Transfection parameters were thereby optimized for a variety of adherent and non-adherent human cancer cell lines resulting in transfection efficiencies of up to 99%; while maintaining high cell viability. Citation Format: Jenny Schroeder, Ludger Altrogge, Elke Lorbach, Srinivasan Kokatam, Sabine Schaepermeier, Meike Weigel, Gina Andretta-Beu, Stefanie Buesch, Tamara Grabeck, Alexandra Krumnow, Sonja Spicker, Sampada Kallol, Preeti Kapoor, Andrea Toell. Efficient transfection of cancer cell lines using the 4D-NucleofectorTM System. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3485. doi:10.1158/1538-7445.AM2014-3485
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