Transfection of hPSC-Cardiomyocytes Using Viafect™ Transfection Reagent

Figures

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  Figure 1. Publications by year in regard to human pluripotent stem cells (hiPSC)s, human pluripotent stem cell-derived cardiomyocytes (hiPSC-CM)s and their transfection [21].

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  Figure 2. Optimisation of hiPSC-CM transfection with Viafect™. To determine the experimental conditions ensuring highest TE, hiPSC-CMs were replated in 96 well plate format and transfected with reporter plasmid pmaxGFP under several combinations of serum content and Viafect:DNA ratios. (a) High content fluorescence imaging enabled determination of transfection efficiency (TE) in hiPSC-CMs; (b) Optimisation of serum content and Viafect to DNA ratio in 96 well plate format revealed that (c) transfection with 6:1 Viafect:DNA ratio in medium containing 10% foetal bovine serum (FBS) achieves >95% TE; (d) Sustained expression of a lncRNA in hiPSC-cardiomyocytes. The results were normalised to an untransfected control. n = 3 biological replicates (4 days and 7 days), n = 1 (14 days). Scale bars in (a) = 1 mm, (b) = 10 mm.

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  Figure 3. Seeding density affects transfection efficiency. hiPSC-CMs were seeded at a range of densities, from 1 × 104 to 5 × 104 per well, then transfected with 6:1 Viafect to DNA with 10% serum supplementation, 3 days post replating. Live images taken 3 days post transfection (upper panel GFP only, lower panel GFP and brightfield overlay). Scale bar = 1 mm.

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  Figure 4. hESC-CMs can be successfully transfected with Viafect. Live images showing brightfield and GFP fluorescence. hESC-CMs were transfected with the reporter plasmid pmaxGFP, using the same parameters of serum, content, reagent: DNA ratio, seeding density and timing determined by the hiPSC line. Scale bar = 1 mm.