Guide to the heart: Differentiation of human pluripotent stem cells towards multiple cardiac subtypes

Schwach, V. (2020)

Doctoral Thesis

General aim of this thesis, entitled Guide to the heart, was to explore the generation of multiple human pluripotent stem cell (hPSC)-derived cardiac subtypes and their application for selective pharmacology, understanding human cardiac development and cardiac repair.

Approaches for the differentiation of hPSCs to cardiomyocytes (CMs) followed by purification from heterogeneous cultures are described. To generate subtype specific CMs, a protocol for the derivation and characterization of hPSC-derived CMs with atrial identity was developed. HPSC-derived atrial CMs have proven successful as pre-clinical pharmacological tool. The development of a human atrial reporter by CRISPR/Cas9-mediated knockin of red fluorescent mCherry into the genomic locus of atrial-enriched COUP-TFII allowed the selection of atrial and ventricular CMs. In addition, we evaluated the importance of COUP-TFII for atrial differentiation of hPSC and identified that COUP-TFII is dispensable for atrial differentiation of hPSCs. Importantly, hPSC-derived cardiac progenitors (CPCs) alleviated ventricular remodeling and fibrosis after transplantation to the heart in an acute myocardial infarction model in mice. This thesis ends with a review of the native cardiac environment during development, as well as the adult heart in health and disease. This was used to describe current knowledge regarding extracellular matrix preferences for engineering cardiac tissues from hPSC-CMs.

• Human pluripotent stem cells
• Cardiomyocytes
• Cardiac differentiation
• Atrial specification
• COUP-TF
• Selective pharmacology
• CRISPR-Cas9
• Cardiac progenitors
• Cardiac repair
• Cardiac extracellular matrix

• All authors: Schwach, V.
• Supervisor: Mummery, C.; Passier, R.
• Co-supervisor: Salvatori, D.
• Committee: Goumans, M.J.; Sluiter, J.; Velden, J. van der; Pijnappels, D.
• Qualification: Doctor (dr.)
• Awarding Institution: Leiden University of Medical Center [LUMC], Leiden University
• Date: 2020-01-15
• ISBN (print): 9789463239059