Engineering of Ferritin Overexpression in Human Induced Pluripotent Stem Cell Derived Cardiomyocytes

Heart injury and disease following an instance of infarction is one of the most common causes of mortality across the world. Fortunately, experiments involving the transplantation of human induced pluripotent stem cell-derived cardiomyocytes have recently indicated that this therapy has the potential to remuscularize and therefore heal damaged cardiac tissue while improving overall cardiac function. While this may be possible, further advancements in studies have been hindered due to a lack of understanding of the long term effects of this technique. The hypothesis to be tested is that ferritin overexpression in hiPSC-CMs will allow post transplant tracking of cells using magnetic resonance imaging due to the ability to function as a contrast agent. Two different approaches to increase cellular levels of ferritin were attempted. These include electroporation of a plasmid expressing ferritin and transfection of a CRISPR activation plasmid. Transfection efficiency in hiPSC-CMs was evaluated by testing cell viability and concentration of stored iron chloride. Electroporation proved to be unsuccessful by weak signals from the GFP reporter protein. However, overexpression of ferritin in hiPSC-CMs following CRISPR activation plasmid transfection demonstrated high cell viability, an appropriate detection of the presence of iron chloride, and minimal toxicity. The use of iron chloride in this study was also shown to have minimal to no toxic effects. Overall, cells were successfully engineered to overexpress ferritin and will be undergoing MRI testing to determine the efficacy of tracking using this method.