Modelling cardiac fibrosis using three-dimensional cardiac microtissues derived from human embryonic stem cells

Abstract

Background Cardiac fibrosis is the most common pathway of many cardiac diseases. To date, there has been no suitable in vitro cardiac fibrosis model that could sufficiently mimic the complex environment of the human heart. Here, a three-dimensional (3D) cardiac sphere platform of contractile cardiac microtissue, composed of human embryonic stem cell (hESC)-derived cardiomyocytes (CMs) and mesenchymal stem cells (MSCs), is presented to better recapitulate the human heart. Results We hypothesized that MSCs would develop an in vitro fibrotic reaction in response to treatment with transforming growth factor-β1 (TGF-β1), a primary inducer of cardiac fibrosis. The addition of MSCs improved sarcomeric organization, electrophysiological properties, and the expression of cardiac-specific genes, suggesting their physiological relevance in the generation of human cardiac microtissue model in vitro. MSCs could also generate fibroblasts within 3D cardiac microtissues and, subsequently, these fibroblasts were transdifferentiated into myofibroblasts by the exogenous addition of TGF-β1. Cardiac microtissues displayed fibrotic features such as the deposition of collagen, the presence of numerous apoptotic CMs and the dissolution of mitochondrial networks. Furthermore, treatment with pro-fibrotic substances demonstrated that this model could reproduce key molecular and cellular fibrotic events. Conclusions This highlights the potential of our 3D cardiac microtissues as a valuable tool for manifesting and evaluating the pro-fibrotic effects of various agents, thereby representing an important step forward towards an in vitro system for the prediction of drug-induced cardiac fibrosis and the study of the pathological changes in human cardiac fibrosis. Electronic supplementary material The online version of this article (10.1186/s13036-019-0139-6) contains supplementary material, which is available to authorized users.

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

“…Alternatively, a trans-well culture with a physical separation of two cell types allows for the study of paracrine interactions but does not include direct cell-cell signaling or the presence of native ECM. CF culture in scaffold-free, multicellular aggregates does not lead to significant proliferation of primary CF, even on the surface of the spheroid, unless stimulated by growthor other profibrotic factors [(Desroches et al, 2012;Lee et al, 2019), and own observations]. Therefore, 3D co-culture offers an option to study myocardial cells with different proliferative potential and their interactions in vitro, at baseline and in stress conditions.…”

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

“…In our model, TGF-β1 induced fibrotic phenotype in cardiac microtissues, as indicated by elevated levels of fibrotic genes, secretion of procollagen I and fCFs proliferation. Previously, exogenous TGF-β1 has been shown to induce fibrosis in rat [17,18] or human [19] cardiac microtissue models. Furthermore, fibrotic phenotypes in cardiac microtissues or in biowire were also produced by collagen/fibroblast enrichment [20,21] or by constitutive activation of profibrotic pathways [22].…”

Development of quantitative direct prediction algorithm for the human target organ similarity of human pluripotent stem cell-derived organoids and cells

“…Moreover, transcriptome analysis showed higher sensitivity and reproducibility than proteome analysis. However, although transcriptome analysis can predict tissue-specific features, RNA-seq data do not reflect 100% of cell functions because total RNA is not translated as proteins 28 Researchers generally performed FACS analysis with CM-specific markers (cTnT) to evaluate the CM differentiation rate derived from hPSCs 29 . In this study, the CM control and CM+BMP4 groups showed 29% and 92% in FACS analysis, but HtGEP analysis showed 34.3% and 83.4% (Fig.…”