TABLE 2.

Overview of iPSC-based 3D cell culture approaches.

Type	Bio-functional properties	Advantage	Disadvantage	References
Spheroids	Self-assembling multicellular aggregates.	Scaffold-free technology without the use of any exogenous support. Composition with different cell-type populations. Manipulation by pipetting and sedimentation. Size allows the miniaturized multi-well formats compatible with plate readers	Small size and heterogeneous composition of multicellular aggregates. No direct measurements of electrophysiological properties. Non-linear cell alignment. Number of cells: detrimental for the cellular viability.	Polonchuk et al., 2017; Tan et al., 2017; Correia et al., 2018; Hoang et al., 2018; Kim T. Y. et al., 2018; Mattapally et al., 2018; Yan et al., 2019
Organoids	Self-assembling organ-like tissue.	Composition with different cell-type populations. Mimics thicker tissues. High level of tissue organization. Size allows the miniaturized multi-well formats compatible with plate readers.	Stochastic events for self-assembly: heterogeneity in shape, size and cell composition. Size of multicellular aggregates. Number of cells: detrimental for the cellular viability.	Kusuma et al., 2013; Lancaster et al., 2013; Orlova et al., 2014; Chan et al., 2015; Eder et al., 2016; Chen Y.-W. et al., 2017; Richards et al., 2017; Mills et al., 2019; Wimmer et al., 2019
Cell sheet cardiac patches	Transplantable 3D-constructs.	Layering of CMs forming a tissue-like structure. Easy to produce and to manipulate. Precise control of cell sheet shape and structure.	Number of cell layers: possible decrease in oxygen and nutrients supply to cells; necrosis of the cells sheets after implantation.	Shimizu et al., 2006; Stevens et al., 2009; Gaetani et al., 2012; Vallée et al., 2012; Wendel et al., 2015; Masumoto et al., 2016; Ishigami et al., 2018; Noor et al., 2019; Park et al., 2019
Engineered cardiac tissue models	3D heart-like constructs with higher level of biological complexity.	Composition with different cell-type populations. Linear cells alignment. Improved maturation of hIPSC-CMs. Direct measurements of electrical activity of the cells.	Coupling with the host tissue. Vascularization. Biocompatibility of the materials.	Tulloch et al., 2011; Deng et al., 2015; Mannhardt et al., 2016; Masumoto et al., 2016; Sugiura et al., 2016; Weinberger et al., 2016; Hernandez et al., 2018; Maiullari et al., 2018; Ronaldson-Bouchard et al., 2018; Lee et al., 2019; Redd et al., 2019; Traverse et al., 2019; Tsui et al., 2019; Roshanbinfar et al., 2020