Functional arrays of human pluripotent stem cell-derived cardiac microtissues

Abstract

<jats:title>ABSTRACT</jats:title><jats:p>To accelerate the cardiac drug discovery pipeline, we set out to develop a platform that would be amenable to standard multiwell-plate manipulations and be capable of quantifying tissue-level functions such as contractile force. We report a 96-well-based array of 3D human pluripotent stem cell (hPSC)-derived cardiac microtissues - termed Cardiac MicroRings (CaMiRi) - in custom printed multiwell plates capable of contractile force measurement. Within each well, two elastomeric microcantilevers are situated above a ramp. The wells are seeded with cell-laden collagen which, in response to the slope of the ramp, self-organizes around tip-gated microcantilevers to form contracting CaMiRi. The contractile force exerted by the CaMiRi is measured and calculated using the deflection of the cantilevers. Platform responses were robust and comparable across wells and we used it to determine an optimal tissue formulation. We validated contractile force response of CaMiRi using selected cardiotropic compounds with known effects. Additionally, we developed automated protocols for CaMiRi seeding, image acquisition, and analysis to enable measurement of contractile force with increased throughput. The unique tissue fabrication properties of the platform, and the consequent effects on tissue function, were demonstrated upon adding hPSC-derived epicardial cells to the system. This platform represents an open-source contractile force screening system useful for drug screening and tissue engineering applications.</jats:p>

Circulating re-entrant waves promote maturation of hiPSC-derived cardiomyocytes in self-organized tissue ring

“…To achieve higher degrees of hiPSC-derived cardiomyocytes maturation and function, a variety of methods have been developed, including dynamic culture 5 , three-dimensional (3D) engineered heart tissue [6][7][8] , 3D printing 9 , addition of factors 10 , extracellular matrix 10,11 , and external stimulation. Electrical stimulation, especially high-frequency pacing, has long been suggested as an effective method for maturing muscle cells 3,[12][13][14][15] , which upregulates cardiac markers 13,14,16 , enhances Ca 2+ -handling properties 3 , and promotes cardiomyocyte alignment 17 .…”

Organotypic and Microphysiological Human Tissue Models for Drug Discovery and Development—Current State-of-the-Art and Future Perspectives

“…Although high-throughput drug screening using EHTs was initially challenging because of their size, several approaches achieved suitably small tissues (Hansen et al, 2010;Boudou et al, 2012), and EHT constructs have by now been established in 96-well format (Mills et al, 2017;Thavandiran et al, 2020). Known compounds such as quinidine, chromanol, erythromycin, and doxorubicin have been tested on EHTs in dose-dependent manner (Hansen et al, 2010).…”

From Human Pluripotent Stem Cells to 3D Cardiac Microtissues: Progress, Applications and Challenges

“…In both models, different cardiac cells, including ECs and CFs, which can be hPSC-derived or primary cultured cells, are combined with hPSC-derived CMs at a specific ratio. In the case of the EHT models, the composition of the hydrogel and the concentration of the ECM used are also important parameters to ensure tissue structure and functionality [87].…”

“…The EHT models described in the literature are composed of hPSC-derived CMs alone [88][89][90][91] or in combination with primary fibroblast/stromal cells [92][93][94] and endothelial cells (HUVECs) [95] normally embedded in an hydrogel-based matrix that is then shaped according to a specific format. One of the models that has been described is the ring-shaped EHT, in which the mixture is pipetted into circular casting molds, where the tissue condenses, and is then placed around passive-flexible holders [87,92,96]. A different strategy relies on the development of elliptic shaped or strip-like cardiac tissues, which are anchored and stretched between two flexible pillars [89,90,93,95,97,98].…”

“…In the case of EHT, the adaptation to a HTS format has been more challenging, mainly due to the size of the construct that should be compatible with a multi-well platform, to maximize the throughput of the system, and also due to the number of hPSC-derived cardiac cells needed for each cardiac MT. To overcome those limitations, small size EHT platforms have been described in the literature, including the Heart-Dyno platform [98,102], the Cardiac MicroRings (CaMiRi) [87] and the µTUG arrays [95]. Some platforms are now commercially available, such as in the case of the Biowire TM II platform (TARA Biosystems) [94,117] and the cardiac tissue strip model (Novoheart TM ) [127,128] (see Table 2).…”