Date of Award

7-2016

Document Type

Thesis

Degree Name

Master of Science (MS)

Legacy Department

Bioengineering

Committee Member

Dr. Agneta Simionescu, Committee Chair

Committee Member

Dr. Dan Simionescu

Committee Member

Dr. Kevin Champaigne

Abstract

Heart disease includes an array of cardiovascular diseases causing 25% of all deaths annually in the United States. Patients who live with heart disease experience significantly decreased quality of life. While current treatments on the market aim at improving heart function and cardiovascular efficiency, none are able to restore functionality to native levels. Tissue engineered approaches are becoming increasingly viewed as the definitive treatment to increasing longevity and quality of life of patients affected by heart disease. While stem cell therapies have immense potential, clinical application is still largely unsuccessful in generating healthy myocardium in patients. This study aims to differentiate and characterize human embryonic, induced pluripotent, and adipose derived stem cells into cardiomyocytes to engineer an electrically conductive construct for heart disease patients. Action potentials of embryonic, and induced pluripotent derived cardiomyocytes were measured using Nanion’s automated patch clamp system in order to attain baseline measurements. Current and voltage clamp configurations were used under physiological conditions and electrical signals were sampled every 15 seconds. In addition, two and three dimensional co-culture studies were performed using adipose-derived and embryonic-derived cardiomyocytes. The cells were seeded within an agarose gel and on 6-well plates at a 2:1 ratio. Samples were then measured using the automated patch clamp system after seven days of culture and stained for connexin-43 and desmin using immunofluorescence. Lastly, multielectrode array measurements readings were performed using embryonic derived cardiomyocytes to verify the patch clamp data. Results showed differentiated induced pluripotent stem cells produced action potentials almost identical to ventricular cardiomyocytes. No action potential readings were able to be recorded for the three dimensional cultures, however physiologically relevant action potentials were measured in the 2D co-culture groups. Additionally, the 3D co-culture samples all stained positive for connexin-43 and desmin verifying that the differentiated cells possessed unique cardiomyocyte markers. Future studies should devote more effort into differentiating successful cardiac pacemaker cells from adipose derived stem cells.

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