Creation of αMHC-GCaMP8 plasmid to monitor the differentiation of human iPSCs into cardiomyocytes
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Due to the poor regenerative potential of adult heart tissue, heart disease is a major cause of death worldwide. With developments in cell biology, researchers have gravitated towards investigating the applications of stem cells to aid in cardiac regeneration through injection of cardiac progenitors into the site of infarction. This method has led to promising results in mammalian studies but to bolster current cardiac research, research tools are required to observe the physiology and quantify the success of differentiation in ES derived cardiac cells. I have attempted to create one such research tool by creating a visual and quantifiable detection method for the differentiation of human iPS cells into beating cardiomyocytes through my creation of an αMHC-GCaMP8 insertion plasmid. Transforming this plasmid into iPSCs will allow GCaMP8 to fluorescently monitor Calcium levels in differentiated cardiomyocytes, taking advantage of the physiological Calcium fluctuations associated with muscle contractions. Though I was unable to confirm creation of this plasmid due to time constraints and unexpected errors, my work can still be finished or expanded upon transforming cells with my plasmid and creating cells with countless applications both in differentiation analysis, disease modeling, and tissue engineering.