Abstract
Magnetic nanoparticle-based gene transfection has been shown to be an effective, non-viral technique for delivery of both plasmid DNA and siRNA into cells in culture. It has several advantages over other non-viral delivery techniques, such as short transfection times and high cell viability. These advantages have been demonstrated in a number of primary cells and cell lines. Here we report that oscillating magnet array-based nanomagnetic transfection significantly improves transfection efficiency in both human prenatal cardiac progenitor cells and adult cardiomyocytes when compared to static magnetofection, cationic lipid reagents and electroporation, while maintaining high cell viability. In addition, transfection of adult cardiomyocytes was improved further by seeding the cells onto Collagen I-coated plates, with transfection efficiencies of up to 49% compared to 24% with lipid reagents and 19% with electroporation. These results demonstrate that oscillating nanomagnetic transfection far outperforms other non-viral transfection techniques in these important cells.
Highlights
The discovery, isolation and differentiation of human cardiac progenitor cells from the epicardium of the heart has given scientists and clinicians alike a tangible opportunity to investigate cardiovascular diseases as well as other issues concerning tissue regeneration [1,2,3,4]
Adhesion Assay As human adult cardiomyocytes are a semi-adherent cell type, adhesion studies were performed. 96 well tissue culture plates (Corning, New York, USA) were coated with foetal bovine serum (FBS: 10, 5, 2.5, and 1.25%) (Lonza, Cologne, Germany), polyethyleneimine (PEI: 10, 5, 2.5, 1.25, 0.6 and 0.3 mg/ml) (Sigma, Dorset, UK), gelatin (0.1, 0.05 and 0.025% w/v) (Sigma, Dorset, UK) and rat tail collagen I (1, 0.5, 0.25 and 0.125 mg/ml) (Sigma, Dorset, UK), made up in phosphate buffer saline (PBS) and incubated overnight at 4uC
In prenatal human cardiac progenitor cells approximately 18.665.2% transfection efficiency was observed when the complexes were subjected to an NdFeB magnet array oscillating at a frequency of 2 Hz with a displacement of 0.2 mm (Figure 2)
Summary
The discovery, isolation and differentiation of human cardiac progenitor cells from the epicardium of the heart has given scientists and clinicians alike a tangible opportunity to investigate cardiovascular diseases as well as other issues concerning tissue regeneration [1,2,3,4]. The isolation of primary adult human cardiomyocytes has made it possible to formulate in vitro models to understand the human heart and cardiac diseases [7], identify the different cardiomyocytes present [8], and study cardiomyocyte differentiation to address tissue regeneration [9]. Despite these advances, successes in cardiovascular gene therapy still remains elusive and non-viral transfection of cardiomyocytes suffers from poor efficiency and relatively low cell viability. Most gene delivery approaches are not been widely applicable due to low transfection efficiency or the lack of suitable vectors, target specificity, or safety issues arising from translating the technique into humans [11]
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