Efficient transfection of primary cells relevant for cardiovascular research by nucleofection.
Cell types that are important for cardiovascular research, e.g., cardiomyocytes, endothelial cells, or adult stem cells, are often hard to isolate, culture, and transfect. Low-transfection efficiencies are a major limitation because, in many cases, results achieved with surrogate model cell lines, if any at all are available for the primary cell type of interest, do not reflect the situation in the primary cell. We have demonstrated that unprecedented transfection results are achieved with primary cells when novel electroporation conditions are combined with a treatment of the cells in specific solutions that help stabilize the cells in the electrical field. This led to the development of the new proprietary transfection technology nucleofection. Nucleofection has proved to be successfully applicable to a variety of primary cells and other hard-to-transfect cell lines, and, thus, opens unique perspectives for novel experimental setups as therapeutic strategies. Herein we present protocols for the efficient nucleofection of human umbilical vein endothelial cells, human coronary artery endothelial cells, smooth muscle cells (e.g., pig vascular smooth muscle cells), neonatal rat cardiomyocytes, and human mesenchymal stem cells and depict some results obtained with such transfected cells.
Thiel
Corinna
C
Amaxa Biosystems, Cologne, Germany.
Nix
Michael
M
D000818:Animals; D018929:Cell Culture Techniques; D003331:Coronary Vessels / Q000166:cytology; D018274:Electroporation / Q000379:methods; D042783:Endothelial Cells; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans / Q000235:genetics; D059630:Mesenchymal Stem Cells; D008856:Microscopy, Fluorescence; D032383:Myocytes, Cardiac; D032389:Myocytes, Smooth Muscle; D051381:Rats / Q000235:genetics; D014162:Transfection / Q000379:methods*; D014471:Umbilical Veins / Q000166:cytology