3D Cardiac Cell Culture: A Critical Review of Current Technologies and Applications

Abstract

Three-dimensional (3D) cell culture is often mentioned in the context of regenerative medicine, for example, for the replacement of ischemic myocardium with tissue-engineered muscle constructs. Additionally, 3D cell culture is used, although less commonly, in basic research, toxicology, and drug development. These applications have recently benefited from innovations in stem cell technologies allowing the mass-production of hiPSC-derived cardiomyocytes or other cardiovascular cells, and from new culturing methods including organ-on-chip and bioprinting technologies. On the analysis side, improved sensors, computer-assisted image analysis, and data collection techniques have lowered the bar for switching to 3D cell culture models. Nevertheless, 3D cell culture is not as widespread or standardized as traditional cell culture methods using monolayers of cells on flat surfaces. The many possibilities of 3D cell culture, but also its limitations, drawbacks and methodological pitfalls, are less well-known. This article reviews currently used cardiovascular 3D cell culture production methods and analysis techniques for the investigation of cardiotoxicity, in drug development and for disease modeling.

Keywords: 3D cell culture; cardiomyocyte; engineered heart tissue; high content screening; induced pluripotent stem cells; scaffold; spheroid.

Figure 1

Overview of different types of EHTs. Reprinted with permission from Weinberger et al. (11). (A) Plane EHT on Velcro-covered rods (5), (B) fabrication of ring-shaped EHTs (12), (C) fibrin-based mini-EHT on polydimethylsiloxane (PDMS) racks (13), (D) cardiac micro tissues (CMT) on fluorescent pillars (14), (E) cardiobundles on a PDMS frame (15), (F) micro heart muscle (16), (G) cardiac biowires (17), (H) cardiac patch (18).

Figure 2

Overview of different spheroid production methods. (A) Hanging drops (InSphero, GravityPlus), (B) U-shaped multi-well plate with non-adhesive coating (Greiner bio-one, Cellstar), (C) view inside a U-shaped well at day 2 of culture, (D) Spheroid made in hanging drop for 3 days was transferred to a non-adhesive multi-well plate for further culture (InSphero, GravityTrap), (E) Silicone micro-mold (3D Petri-dish, microtissues.com) for making an agarose cast for 81 spheroids, (F) Cardiac spheroids reside inside the 3D petri-dish agarose cast submerged in medium 4 days after seeding of the cell solution, (G) Small cardiac spheroids forming 3 days after seeding in a micro-patterned multi-well plate with 12 × 750 microwells (Kugelmeiers, Sphericalplate 5D). All photos by the author.