Human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) have the potential to be an important tool for cardiovascular disease modeling, pre-clinical cardiotoxicity evaluation, and drug discovery. However, detailed studies of their excitation-contraction (EC) coupling are limited by the lack of appropriate methodology. Here, I set out to investigate the EC coupling of normal and diseased hiPSC-CMs and compare the results to that of adult ventricular CMs (i.e., rabbit and mouse) under identical experimental conditions. I found that hiPSC-CMs display relatively mature EC coupling properties (i.e., electrophysiology, Ca handling and contractility). To assess their contractile properties I have developed a novel culture method that enables robust contractile measurements of single hiPSC-CMs. I discovered that hiPSC-CMs display contractile properties comparable to that of adult rabbit CMs, including comparable contraction kinetics. Moreover, EC coupling properties were comparable across hiPSC-CM lines generated at different institutions and post recovery from cryopreservation. Using disease specific hiPSC-CMs I revealed that HCM MYH7 R633H hiPSC-CMs display contractile abnormalities, and I provided evidence supporting an enhanced myofilament Ca sensitivity mechanism. Furthermore, I demonstrated hiPSC-CMs display a robust response to pharmacological stimuli including the myofilament Ca sensitizer EMD57033. These findings will aid functional studies of disease specific hiPSC-CMs as well as the effects of novel and known pharmacological agents.
| Identifer | oai:union.ndltd.org:VANDERBILT/oai:VANDERBILTETD:etd-11182015-125322 |
| Date | 18 November 2015 |
| Creators | Feaster, Tromondae K |
| Contributors | Charles C. Hong, Joey V. Barnett, Björn C. Knollmann, Dan M. Roden, H. Scott Baldwin |
| Publisher | VANDERBILT |
| Source Sets | Vanderbilt University Theses |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.library.vanderbilt.edu/available/etd-11182015-125322/ |
| Rights | unrestricted, I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to Vanderbilt University or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. |
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