The interplay between hESC-derived heart progenitors and the extracellular matrix towards developing regenerative medicine treatments and investigative tools for congenital heart disease

  • Tuva R Wegnelius Jarlstedt

Student thesis: Doctoral ThesisDoctor of Philosophy (PhD)

Abstract

Background: Congenital heart disease (CHD) is currently the largest cause of birth defects. Generation of new investigative tools and regenerative medicine treatments for CHD is therefore needed. Here the interplay between hESC-derived heart progenitors and the extra cellular matrix (ECM) is studied to develop regenerative medicine treatment and tools for CHD.

Methodology: 1. hESC-derived cardiomyocytes, epicardium cells, cardiac fibroblast, and endothelial cells were generated in vitro and characterised using established protocols for the development of regenerative medicine approaches to treat CHD. 2. To generate a transplantable multilineage heart patch using a decellularized ECM, the hESC-derived cardiac and cardiovascular cells (70% CM, 15% CFs, and 15% EC) were seeded onto a ProxiCor matrix in combination. The properties of the multilineage patch were characterised using mechanical testing, histology, ICC and electrophysiology testing. 3. To examine the in-house established ECM component rAgrin effect on cardiomyocyte proliferation, hESC-CMs were treated with 20ng/ml, 100ng/ml, and 500ng/ml rAgrin and the cells proliferation measure using cell-cycle markers AURKB and PH3 (n=8). 4. Generated knockout cell line using CRISPR gene editing technology as a tool to study the effects of known CHD mutation on lineage development and the interaction of CHD-disease model with the ECM.

Results: The multilineage cardiac patch was successfully generated and showed lower Youngs Modulus for ProxiCor seeded with multiple cardiac cells (n=3, p<0.05), indicating a potential remodelling and relaxation in the matrix. Contracting hESC-CMs were also recorded on the ProxiCor 7 days after seeding (n=3). The hESC-CMs treated with rAgrin showed an increased expression of cell-cycle markers AURKB and PH3 compared to the untreated control (n=3, p<0.05).

Conclusion: It can be concluded that a multilineage heart patch were generated using established hESC-derived cardiac and cardiovascular cells, taking us closer to the possibility of a hESC-derived multilineage patch for CHD treatment. It can also be concluded that rAgrin effects cardiomyocyte proliferation.
Date of Award18 Jun 2024
Original languageEnglish
Awarding Institution
  • University of Bristol
SupervisorMassimo Caputo (Supervisor), Paolo R Madeddu (Supervisor) & Victoria Mascetti (Supervisor)

Cite this

'