Cardiomyocyte dedifferentiation and remodeling in 3D scaffolds to generate the cellular diversity of engineering cardiac tissues

The use of engineered cardiac tissues (ECTs) is a new strategy for the repair and replacement of cardiac tissues in patients with myocardial infarction, particularly at late stages. However, the mechanisms underlying the development of ECTs, including cell-scaffold interactions, are not fully understood, although they are closely related to their therapeutic effect. In the present study, we aimed to determine the cellular fate of cardiomyocytes in a 3D scaffold microenvironment, as well as their role in generating the cellular diversity of ECTs by single-cell sequencing analysis. Consistent with the observed plasticity of cardiomyocytes during cardiac regeneration, cardiomyocytes in 3D scaffolds appeared to dedifferentiate, showing an initial loss of normal cytoskeleton organization in the adaptive response to the new scaffold microenvironment. Cardiomyocytes undergoing this process regained their proliferation potential and gradually developed into myocardial cells at different developmental stages, generating heterogeneous regenerative ECTs. To better characterize the remodeled ECTs, high-throughput single-cell sequencing was performed. The ECTs contained a wide diversity of cells related to endogenous classes in the heart, including myocardial cells at different developmental stages and different kinds of interstitial cells. Non-cardiac cells seemed to play important roles in cardiac reconstruction, especially Cajal-like interstitial cells and macrophages. Altogether, our results showed for the first time that cells underwent adaptive dedifferentiation for survival in a 3D scaffold microenvironment to generate heterogeneous tissues. These findings provide an important basis for an improved understanding of the development and assembly of engineered tissues.