Decellularized muscle‐derived hydrogels support in vitro cardiac microtissue fabrication

Abstract

Cardiovascular research has considerably benefited from in vitro models of cardiac tissue. Two important elements of these constructs, cardiac cells and the extracellular matrix (ECM), play essential roles that mimic the structural and functional aspects of myocardium. Here, we compared decellularized ECM from cardiac muscle (D-CM), skeletal muscle (D-SM), aorta (D-Ao), liver (D-Liv), small intestine submucosa (D-SIS), and human umbilical cord (D-hUC) in terms of their biocompatibility and potential for differentiation of human embryonic stem cell-derived cardiac progenitor cells (hESCderived CPCs) to cardiovascular lineage cells. The decellularization procedure successfully removed resident cells of the tissues but preserved ECM components such as laminin and fibronectin, which was identified by histological studies of decellularized tissue (D-tissues) and immunostaining. Encapsulation of hESC-derived CPCs and human umbilical vein endothelial cells within hydrogels that were obtained from all decellularized tissues did not induce cytotoxicity after 10 days of culture. Upregulation of cardiac specific genes, cTNT and αMHC, as well as the presence of cTNT + cardiomyocytes were also observed in CPCs cultured on D-CM, D-SM, D-Liv, and D-SIS, which showed their support for cardiogenic differentiation. However, D-CM provided substantially higher expression of cardiac markers compared to the other D-tissues. The endothelial and smooth muscle specific genes, CD31 and PDGFRα, were upregulated in cells cultured on D-Ao and D-hUC, which reflected their support for vascular lineage cell differentiation. In conclusion, it might be imperative to use decellularized tissue of muscle origins in combination with naturally derived vascular tissues to generate in vitro vascularized human cardiac microtissues. K E Y W O R D S cardiac progenitor cells, extracellular matrix, hydrogel, tissue engineering 1 | INTRODUCTION Regenerative therapy for cardiac ischemic diseases is a developing scientific field that has attracted considerable attention. In particular, tissue engineering has emerged as a very promising supplement to favorable cell therapy approaches (Saludas, Pascual-Gil, Prosper, Garbayo, & Blanco-Prieto, 2017). Acellular therapies based on tissue engineering have shown substantial outcomes (Liberski, Latif, Raynaud, Bollensdorff, & Yacoub, 2016; Traverse et al., 2019). Among proposed biomaterials, the extracellular matrix (ECM) provides cells with environments that have the highest similarity to their native niche (Yi, Ding, Gong, & Gu, 2017), resulting in similar biochemical profiles and growth factor reserves that help to mimic the tissue microenvironment and support function (Freedman et al., 2015;

Heart Repair Induced by Cardiac Progenitor Cell Delivery within Polypyrrole-Loaded Cardiogel Post-ischemia

“…Sheep hearts were provided from a local slaughterhouse. Decellularization was performed as described previously . The full description of the decellularization process as well as its characterization by histological studies and DNA quantification can be found in the Supporting Information.…”

Recent advances in soluble decellularized extracellular matrix for heart tissue engineering and organ modeling

“…16 In addition to primary cardiac ECM (myocardium and pericardium), secondary sources such as small intestine submucosa, omentum, placenta, liver, lung, skin, and urinary bladder matrix have also been studied as potential dECM sources for cardiac tissue engineering applications. 61,85,[123][124][125][126][127][128][129] Although primary sources offer cardiac-specific biochemical cues and structural composition, facile process, ease of obtaining, and improved reproducibility are the main reasons behind using secondary dECM sources for cardiac regeneration. 130 Several studies have been conducted to compare the characteristics of various dECM sources with primary cardiac dECM.…”

Decellularized Extracellular Matrices and Cardiac Differentiation: Study on Human Amniotic Fluid-Stem Cells

“…The realization of an appropriate scaffold for the cardiac tissue engineering presents unique challenges, since native CMs are continuously exposed to cyclic mechanical (stress and strain) and electrical stimuli that must be properly transmitted to neighboring cells. In addition to these peculiar characteristics, tissue scaffolds for heart regeneration must also comply with the general requirements for all tissue scaffolds, such as degradation time, cell–scaffold interactions, vascularization/nutrient delivery, and implantation method [ 12 , 13 ]. Among the biological scaffolds, xenogenic, decellularized extracellular matrix (dECM) recently aroused a great deal of interest [ 3 , 14 , 15 , 16 , 17 , 18 , 19 ] with some matrices already being used in clinical practice; in fact, the elimination of cells from the tissues significantly reduces the risk of inflammatory response and immunological rejection, while preserving the natural three-dimensional structure of the ECM.…”