Authors

N T Vo¹; ²; K Raniga¹; ²; A Nasir¹; ²; D Mosqueira¹; ²; H Hall³; D Pavlovic³; C Denning¹; ²; M Haddrick⁴; 
¹ The University of Nottingham, UK;  ² The University of Nottingham, UK;  ³ University of Birmingham, UK;  ⁴ Medicine Discovery Catapult, UK

Discussion

Cardiac fibrosis is hard to treat, with major reliance on developing and using various animal-based in vitro models and species. We evaluated the human pluripotent stem cell (hIPSC) derived lineages to develop an in vitro model of cardiac fibrosis from isogenic genome-edited human induced pluripotent stem cells (hiPSC) harbouring p.R453C-β-myosin heavy chain (β-HCM) mutation underlying hypertrophic cardiomyopathy (HCM), to understand the effect of cell- and mutation-type on cardiac fibrosis.

Isogenic sets of hPSCs were differentiated into cardiomyocytes (CMs), cardiac fibroblasts (CFs) and cardiac endothelial cells (ECs) using our customized serum-free culture protocols. Reproducible and consistently high differentiation efficiency of hIPSC-ECs and hIPSC-CFs without any purification method confirms for the possible replacement of animal serum products in the standard medium system by serum replacement product. We managed to control the activation of MFs from CFs by TGF-β1 and TGF-β inhibitor for fibrosis modelling. Evaluation of contractility phenotype of hIPSC-βMHC-R453C CMs in 2D identified differences caused by the genetic β-HCM mutation. The project will next develop functional assays to establish the specific phenotypic tool kit for both 2D and 3D (spheroid & EHT) tri-linage models during cardiac fibrosis development. This work will benefit the development of improved platforms for drug validation in the cardiovascular field and result in translational applications in future clinical practice. The validation of the humanised platform in serum-free conditions will help to remove the need for animal use in the field.