High throughput strategies for electrophysiological characterisation of cardiomyocytes

by Fitzwilliam Seibertz
Doctoral thesis
Date of Examination:2023-06-08
Date of issue:2023-08-22
Advisor:Prof. Dr. Niels Voigt
Referee:Prof. Dr. Thomas Meyer
Referee:Prof. Dr. Silvio Rizzoli
Referee:Prof. Dr. Christian Griesinger
Referee:Prof. Dr. Tim Salditt
Referee:Prof. Dr. Tobias Moser
Referee:Prof. Dr. Marc Freichel
Persistent Address: http://dx.doi.org/10.53846/goediss-10066

 

 

Files in this item

Name:F Seibertz_PhD_Without CV.pdf
Size:23.7Mb
Format:PDF
Description:Main Article

The following license files are associated with this item:

Abstract

English

Personalised medicine describes the customised application of medical treatment at a patient-specific level. This type of precision medicine stratifies patients based on their specific genotype or phenotype for targeted therapy. For treatment of cardiovascular disorders, this is not yet clinically feasible due to high monetary costs and the slow rate at which molecular disease mechanisms are determined and treatment modalities can be tested. Scalable cellular models such as induced pluripotent stem cell derived cardiomyocyte technology (iPSC-CM) are required for widespread use, along with a concomitant increase in high throughput measurement strategies for robust assessment of cardiac function. This work aims to demonstrate how state-of-the-art high throughput electrophysiological methods can be successfully implemented into basic science research and could aid future implementation strategies for patient-specific care. Herein, three peer-reviewed and published articles deeply analyse cardiomyocyte function using novel high throughput electrophysiological methods. Using specialised voltage and calcium sensitive dyes in single cells, Article I elegantly identifies and targets a novel arrhythmogenic mechanism in iPSC-CM derived from a patient with genetic dilated cardiomyopathy. Such fluorescent dyes provide accurate and non-invasive readouts of cellular membrane voltage and cytosolic calcium concentration, respectively. Article II contains the first known measurements of primary cardiomyocytes using a market- leading high throughput automated patch-clamp (APC) device. Revolutionary in automating the typically complex and experimenter dependent patch-clamp technique, APC is only now beginning to migrate into academic institutions with wider user applications. Article III utilises multiple measurement modalities including APC and in silico techniques to model age-related variability in iPSC-CM technology and assess the suitability of the construct for mechanistic studies and compound screening in the development of personalised medicine paradigms.
Keywords: Automated patch clamp; Cardiac; Electrophysiology; Stem cells
Schlagwörter: Automated patch clamp; Cardiac; Electrophysiology; Stem cells
 
Statistik