Human induced pluripotent stem cells (hiPSCs) hold great value for their ability to produce differentiated cells such as hiPSC-derived cardiomyocytes (hiPSC-CMs) from any individual of interest. For this reason, hiPSC-CMs have found numerous applications in disease modeling and drug development, and are even being investigated as a potential therapeutic modality. However, there are still challenges associated with being able to reliably produce homogeneous populations of the desired cell types at the appropriate developmental stage. Chapter I reviews the types and sources of inter-culture variability and intra-culture heterogeneity in iPSCs and their differentiated cardiomyocyte progeny, including in features associated with ventricular, atrial, and nodal cardiomyocyte subtypes. This chapter also describes a framework for characterizing and modulating cellular properties in accordance with the desired application. Chapter II details the materials and methods used throughout this thesis. Chapter III documents the heterogeneous and dynamic nature of hiPSC-CM electrophysiology and single-cell transcriptomes over time in culture. This chapter reveals the complex nature of hiPSC-CM action potential profiles, which can be influenced by the expression of individual genes and may not be predictive of the expression patterns of canonical cardiomyocyte subtype markers. Chapter IV reveals the impact of non-cardiomyocytes produced in the cardiac differentiation process upon hiPSC-CM electrophysiology, highlighting an important environmental influence upon hiPSC-CM phenotyping. Chapter V summarizes and synthesizes the main findings of these studies, describes a novel model for the development of hiPSC-CM subtype-associated phenotypes in the context of the cellular environment, and proposes future directions for the hiPSC-CM platform based on the conclusions of this thesis.
|Advisor:||Nelson, Timothy J.|
|Commitee:||Ackerman, Michael J., Ekker, Stephen C., Griffiths, Leigh G., Li, Hu, Lu, Lichun|
|School:||College of Medicine - Mayo Clinic|
|Department:||Molecular Pharmacology and Experimental Therapeutics|
|School Location:||United States -- Minnesota|
|Source:||DAI-B 80/11(E), Dissertation Abstracts International|
|Subjects:||Cellular biology, Pharmacology|
|Keywords:||Cardiac, Cell communication, Cell differentiation, Electrophysiology, Gene expression, Induced pluripotent stem cells, Myocytes|
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