The parent grant ?Engineered Human Heart Slice for Testing Drug-Induced Arrhythmia? aims to develop an engineered hear slice (EHS) model that will have the ability to test for drug-induced, cardiac tachyarrhythmias. The cardiomyocytes in the EHS model are obtained from human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). While hiPSC-CMs are a promising cell source for cardiomyocytes, these cells are limited in their maturation state. hiPSC- CMs are currently fetal-like in their development, and require stimulants to achieve an adult-like phenotype. The parent grant focuses on the use of electromechanical, biological, and maturation medias to attain such a phenotype in hiPSC-CMs. This leads to the goal of this supplemental research, which is to apply an alternate stimulant to aid in the maturation of hiPSC-CMs. This stimulant is the secretome from adipocytes (fat generating cells). The adipocytes will serve as a source of fatty acids, which have proven to be influential in the metabolic maturation of cardiomyocytes.
The specific aim of this supplemental work is to investigate the beneficial role of the secretome from adipocytes derived from human Adipose-Derived Stem Cells (hASCs) as a maturation stimulant for hiPSC-CMs. The secretome from adipocytes will be integrated into the hiPSC-CM culture media through the addition of conditioned media or use of transwell inserts. The study will also determine whether hiPSC-CMs sorted for CD36+ (a metabolic maturation marker) are particularly responsive to the secretome. hiPSC-CM maturation will be evaluated through morphological, electrophysiological testing, cell-surface markers, single cell RNA-seq and transcriptional profiling.
Developing a cardiac model capable of heart regeneration, disease modeling, and drug screening has been a challenge in cardiac tissue engineering. When human stem cells are used as the source of the cardiomyocytes, major limitations are associated with the immaturity of the cells. This project aims to improve the maturation of stem cell-derived cardiomyocytes through chemical stimulants secreted by patient-derived fat cells.
