3D Co-culture of hiPSC-Derived Cardiomyocytes With Cardiac Fibroblasts Improves Tissue-Like Features of Cardiac Spheroids

Abstract

Both cardiomyocytes and cardiac fibroblasts (CF) play essential roles in cardiac development, function, and remodeling. Properties of 3D co-cultures are incompletely understood. Hence, 3D co-culture of cardiomyocytes and CF was characterized, and selected features compared with single-type and 2D culture conditions. Methods: Human cardiomyocytes derived from induced-pluripotent stem cells (hiPSC-CMs) were obtained from Cellular Dynamics or Ncardia, and primary human cardiac fibroblasts from ScienCell. Cardiac spheroids were investigated using cryosections and whole-mount confocal microscopy, video motion analysis, scanning-, and transmission-electron microscopy (SEM, TEM), action potential recording, and quantitative PCR (qPCR). Conclusion: We demonstrate that the use of 3D co-culture of hiPSC-CMs and CF is superior over 2D culture conditions for co-culture models and more closely mimicking the native state of the myocardium with relevance to drug development as well as for personalized medicine.

Activated Cardiac Fibroblasts Control Contraction of Human Fibrotic Cardiac Microtissues by a β-Adrenoreceptor-Dependent Mechanism

“…We confirm that CFs significantly increased the integrity and synchronized contractility of human cardiac microtissues. As recently demonstrated, the use of a 4:1 ratio (iCMs:CFs) results in cardiac microtissues with stable phenotype and beating properties for up to one month [16]. Our data further demonstrated that contractility of these cardiac microtissues was substantially affected under profibrotic condition (i.e., in the presence of TGF-β1).…”

“…Using human iPSC-derived cardiomyocytes (iCMs) and human foetal cardiac fibroblasts (fCFs), we developed a high throughput in vitro model of human cardiac tissue [16]. Cardiac microtissues consisting of 5000 cells each were generated by self-assembly ( Figure S1).…”

“…Panel (C) shows quantification of contraction parameters of cardiac microtissues containing aCFs from unaffected hearts (white) or heart failure (HF) patients (grey). Each dot represents average data of one experiment (n = [14][15][16][17][18]. p values were calculated with the Student's t-test.…”

Bi-directional Impacts of Heterotypic Interactions in Engineered 3D Human Cardiac Microtissues Revealed by Single-Cell RNA-Sequencing and Functional Analysis

“…hiPSC-CMs lack a fully mature phenotype, and transcriptional progression throughout differentiation renders them most comparable to early-/mid-gestation fetal CMs 32–34 . Several key environmental parameters have been implicated in promoting the developmental phenotype of hiPSC-CMs, such as three-dimensional culture 35–38 , biochemical cues 39–41 , electromechanical stimulation 37,38,42–45 , and extended culture duration 46–48 . Consistent with previous studies, the cardiac microtissues examined here exhibited overall improvements in calcium handling properties (amplitude, maximum upstroke and downstroke velocities) with prolonged (30 day) culture in the engineered 3D platform.…”

3D bioprinting of high cell-density heterogeneous tissue models through spheroid fusion within self-healing hydrogels

“…In these formats they are promising as drug screening platforms due to their superior predictability and physiological structure and function when compared to traditional monolayer cultures [7][8][9][10] . Indeed, the organotypic cell densities in spheroids enhances the cell-cell and cell-extracellular matrix (ECM) interactions that are required to maintain cellular differentiation and phenotype, whereas these interactions are limited in traditional 2D cultures [11][12][13][14] . High cell-densities are necessary to accurately recapitulate many pathological disease states such as cancer or fibrosis where perturbed cell-cell and cell-ECM interactions are central to disease progression.…”