Scaffold Free Bio-orthogonal Assembly of 3-Dimensional Cardiac Tissue via Cell Surface Engineering

Abstract

There has been tremendous interest in constructing in vitro cardiac tissue for a range of fundamental studies of cardiac development and disease and as a commercial system to evaluate therapeutic drug discovery prioritization and toxicity. Although there has been progress towards studying 2-dimensional cardiac function in vitro, there remain challenging obstacles to generate rapid and efficient scaffold-free 3-dimensional multiple cell type co-culture cardiac tissue models. Herein, we develop a programmed rapid self-assembly strategy to induce specific and stable cell-cell contacts among multiple cell types found in heart tissue to generate 3D tissues through cell-surface engineering based on liposome delivery and fusion to display bio-orthogonal functional groups from cell membranes. We generate, for the first time, a scaffold free and stable self assembled 3 cell line co-culture 3D cardiac tissue model by assembling cardiomyocytes, endothelial cells and cardiac fibroblast cells via a rapid inter-cell click ligation process. We compare and analyze the function of the 3D cardiac tissue chips with 2D co-culture monolayers by assessing cardiac specific markers, electromechanical cell coupling, beating rates and evaluating drug toxicity.

Programming Cell-Cell Interactions through Non-genetic Membrane Engineering

“…Cells bearing the complementary ketone and oxyamine moieties were then linked to one another via a covalent oxime ligation. When applied to multiple cell types, the Yousaf group has been able to form multicellular sheets (Dutta et al, 2011b), spheroids (O’Brien et al, 2015), and microtissues (Rogozhnikov et al, 2016). …”

“…Ultimately, they were able to use this methodology to assemble fibroblasts, cardiomyocytes, and HUVECs into a functional three-dimensional (3D) cardiac tissue that was able to beat spontaneously and synchronously. Importantly, neighboring cells in this tissue both developed physiologic intercellular junctions – including the gap-junction protein connexin 43 – and produced extracellular matrix, suggesting a more accurate recapitulation of the normal cardiac environment in vivo (Rogozhnikov et al, 2016). Furthermore, the induced cell-cell interactions could be reversed by UV irradiation when a photocleavable group was incorporated into the oxyamine component (Luo et al, 2014).…”

“…This is especially true for tissue engineering applications where a key goal is to drive interactions between the numerous cell lineages comprising a functional tissue or organ. Fortunately, many of the non-genetic approaches reviewed here have each been applied to a wide array of cell types – including lymphocytes (Gabrielse et al, 2014), MSCs (Kean et al, 2012), cardiomyocytes (Rogozhnikov et al, 2016), and vascular endothelial cells (Gong et al, 2013) – suggesting that non-genetic alterations relying solely upon membrane modification may be uniquely flexible and thus find use in a variety of applications.…”

Mechanistic Insights of Cells in Porous Scaffolds via Integrated Culture Technologies

“…Notwithstanding its obvious advantages such as highly reproducible and cost-effective, the time-honored 2D (two-dimensional) cell culture is usually considered as a poor proxy of cells in vivo. This is because the 2D culture conditions are far removed from the complexities cells encounter in real-life tissues [8][9][10]. In vitro 3D tissue models thus have been developed to enhance the physiological relevance of experiments in vitro as they can provide D DAVID PUBLISHING more suitable 3D microenvironments for cell growth, differentiation and function [8][9][10][11].…”

“…This is because the 2D culture conditions are far removed from the complexities cells encounter in real-life tissues [8][9][10]. In vitro 3D tissue models thus have been developed to enhance the physiological relevance of experiments in vitro as they can provide D DAVID PUBLISHING more suitable 3D microenvironments for cell growth, differentiation and function [8][9][10][11]. However, the introduction of these in vitro models are a trade-off between the value of obtaining physiologically relevant data against their through-put as they are usually more labour intensive [11].…”