Development of a Rhesus macaque engineered heart muscle model from pluripotent stem cells

by Brian Golat
Doctoral thesis
Date of Examination:2017-05-15
Date of issue:2017-06-13
Advisor:Prof. Dr. Wolfram-Hubertus Zimmermann
Referee:Prof. Dr. Rüdiger Behr
Referee:Prof. Dr. Lutz Walter
Referee:Prof. Dr. Ralf Dressel
Referee:Prof. Dr. Stefan Luther
Referee:Prof. Dr. Steven Johnsen
Persistent Address: http://dx.doi.org/10.53846/goediss-6342

 

 

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Abstract

English

Myocardial infarction is the leading cause of death globally. Remuscularization of the heart with stem cell based treatments is one potential approach to heart repair which is presently being tested. Proof-of-concept studies, mostly in rodent models, have shown promising results. Nevertheless, to be therapeutically relevant it is essential to test evolving therapeutic strategies in large animal models with high predictive value as to clinical outcome. Unfortunately, a lack of pluripotent stem cell models available in most animal species used for late stage preclinical testing creates a major challenge. Non-human primate models, and here in particular macaque models, appear to be an exception with the documentation of successful derivation of stable pluripotent stem cell models by several groups. In this dissertation the following hypotheses were tested: (1) induced pluripotent stem (iPS) cells from Rhesus macaque (Macaca mulatta) can be used to derive cardiomyocytes, (2) Rhesus macaque iPS-derived cardiomyocytes can be used to construct engineered heart muscle (EHM) with contractile properties similar to observations in human EHM, (3) parthenogenetic pluripotent stem cells can be derived from Rhesus macaque unfertilized oocytes as an alternative source of pluripotent stem cells with distinct advantages regarding immunological matching. Two available Rhesus iPS cell lines, either generated by lentiviral or Sendai viral transduction of reprogramming factors, were subjected to directed differentiation using a similar protocol as recently established for human pluripotent stem cells. High cardiomyocyte yield and purity could be achieved. Cardiomyocytes differentiated from both iPS cell lines could be used to generate EHM with similar contractile properties. Evidence for successful induction of parthenogenesis was obtained. Collectively, this dissertation provides the methodological groundwork for the implementation of Rhesus macaque pluripotent stem cell derived EHMs for pivotal late stage preclinical studies of tissue engineered heart failure repair.
Keywords: Rhesus macaque; EHM; engineered heart muscle; stem cells; heart model; tissue engineering
 
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