Advisory Committee Chair
Advisory Committee Members
Eddy Shih-Hsin Yang
Date of Award
Degree Name by School
Doctor of Philosophy (PhD) School of Engineering
Despite major advances in revascularization and drug therapy for post-myocardial infarction (MI) heart failure (HF), heart transplantation remains the only curative therapy to prevent progressive and debilitating disease. Researchers have long sought a replenishable and expandable source of cardiomyocytes (CMs), the primary functional cell in the heart muscle, for a broad array of applications related to the treatment of HF ranging from direct implantation to high throughput drug testing. The lack of cell cycle activity and cell division in adult CMs prevented the development of an expandable primary cell line. However, with the discovery of high efficiency stem cell differentiation protocols and the ability to reprogram somatic cells from any patient into an induced pluripotent stem cell (iPSC) line, a potentially unlimited source of CMs could be available. Broad use of this tool is still limited by several key factors including scalability of the production systems and immaturity of the resultant product. Currently, the widely used differentiation protocols rely on monolayer techniques that require significant manual labor, take up large amounts of incubator space, and are not well adapted to automation. Here, we leverage suspension culture and bioreactor technologies along with newly available culture medias to develop a novel 3D process to expand iPSCs and differentiate them into highly pure cultures of beating CM spheroids. After differentiation and the beginning of beating around day 12, studies have shown that CMs resemble fetal muscle cells and lack the robust electrical and contractile systems needed to function in an adult heart. This creates a challenge for studies requiring an accurate model as well as implantation therapies where functional status can directly play a role in the effectiveness of a treatment. Chapters 2 and 3 of this dissertation explore methods to introduce complexity and induce rapid maturation towards the development of cardiac organoids with physiological functional activity. We also show a method of production for these micro-myocardial tissues that is well adapted for biomanufacturing and scale-up. These advances create a framework for the scalable production of consistent and easily manipulable cardiac organoids with tissue like structures that can be used in a wide array of applications and will allow for development of novel therapies towards the treatment of post-MI HF.
Kahn-Krell, Asher M., "Biomanufacturing of Micro-Myocardial Tissue Through Suspension Differentiation with Improved Yields, Purity, and Quality" (2022). All ETDs from UAB. 483.