Tissue-embedded stretchable nanoelectronics reveal endothelial cell–mediated electrical maturation of human 3D cardiac microtissues

Abstract

Clinical translation of stem cell therapies for heart disease requires electrical integration of transplanted cardiomyocytes. Generation of electrically matured human induced pluripotent stem cell–derived cardiomyocytes (hiPSC-CMs) is critical for electrical integration. Here, we found that hiPSC-derived endothelial cells (hiPSC-ECs) promoted the expression of selected maturation markers in hiPSC-CMs. Using tissue-embedded stretchable mesh nanoelectronics, we achieved a long-term stable map of human three-dimensional (3D) cardiac microtissue electrical activity. The results revealed that hiPSC-ECs accelerated the electrical maturation of hiPSC-CMs in 3D cardiac microtissues. Machine learning–based pseudotime trajectory inference of cardiomyocyte electrical signals further revealed the electrical phenotypic transition path during development. Guided by the electrical recording data, single-cell RNA sequencing identified that hiPSC-ECs promoted cardiomyocyte subpopulations with a more mature phenotype, and multiple ligand-receptor interactions were up-regulated between hiPSC-ECs and hiPSC-CMs, revealing a coordinated multifactorial mechanism of hiPSC-CM electrical maturation. Collectively, these findings show that hiPSC-ECs drive hiPSC-CM electrical maturation via multiple intercellular pathways.

Engineering Programmable Material-To-Cell Pathways Via Synthetic Notch Receptors To Spatially Control Cellular Phenotypes In Multi-Cellular Constructs

“…Our approach may better mimic natural tissue morphogenesis, where multiple cell fates emerge simultaneously from a uniform cell population. Studies have also shown that supporting cells, such as endothelial cells, improve the maturation of human induced pluripotent stem cellderived cardiomyocytes 59,60 , and that co-differentiation of different lineages concurrently more closely recapitulate the conditions occurring during embryonic development 61 . An interesting hypothesis to explore with our technology is whether co-differentiation of supporting cells (e.g., endothelial cells) adjacent to parenchymal cells (e.g., muscle cells) has additional benefits for phenotypic maturity.…”

Heart-on-a-chip systems: disease modeling and drug screening applications

“…Co-culture of CMs with endothelial cells has also been shown to improve expression of cardiac maturation markers, including Cx43, TNNI3, Kir2.1, and CD36, with a 3 : 1 CM : EC ratio being most favorable. 226 Moreover, this ratio showed improved electrophysiological characteristics and sarcomere organization. While 4 genetically distinct subtypes of CM were identified when hiPSC-CMs were cultured alone, hiPSC-CMs cultured with ECs showed an additional subtype that upregulated genes associated with metabolism and cardiac muscle contraction.…”

“…Given that CMs comprise only 30% by number of total cells in the heart, co-culture with other cell types, such as fibroblasts, neural cells, endothelial cells, and immune cells has been shown to promote maturation through intercellular interaction and paracrine signaling. 8,[212][213][214][215][216][217][218][219][220][221][222][223][224][225][226] Co-culture of hESC-CMs with human cardiac fibroblasts (at an optimized 4 : 1 ratio) in a microfluidic chip containing an array of microposts was found to improve tissue maturity and anisotropy after just 2 weeks (Fig. 3c).…”

The emerging role of heart‐on‐a‐chip systems in delineating mechanisms of SARS‐CoV‐2‐induced cardiac dysfunction

“…The heart is one of the most vascularized organs in the body, with an equivalent number of cardiomyocytes and endothelial cells present in the myocardium 57 . Its complex vasculature both maintains heart homeostasis and promotes cardiac maturation under healthy conditions 157 . Moreover, endothelial cells serve as a physical and chemical barrier in the heart, indicating that direct administration of viruses or drugs to the engineered heart tissue may lead to an overestimation of their effects.…”

“… 57 Its complex vasculature both maintains heart homeostasis and promotes cardiac maturation under healthy conditions. 157 Moreover, endothelial cells serve as a physical and chemical barrier in the heart, indicating that direct administration of viruses or drugs to the engineered heart tissue may lead to an overestimation of their effects. In parallel, the immunomodulatory effects of endothelial cells and immune cells have been implicated in many heart dysfunctions.…”