
Recently, methods based on three‐dimensional (3D) cultivation approaches have steadily evolved to overcome the major limitations of conventional adherent monolayer cultivation (2D). These 3D approaches aim to study, reproduce or modify fundamental native features of the heart, such as tissue organization and cardiovascular microenvironment. This review includes 2D monolayer culturing and the 3D in vitro systems of cardiac spheroids, organoids, engineered cardiac microtissue and bioprinting. The ex vivo technique of myocardial slicing is introduced with its basic concepts, advantages, and limitations. Furthermore, recent advances of various new approaches are presented to extend and optimize these in vitro and ex vivo systems.
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