Fabrication of Multi-cellular Spheroids for Demonstrating the Intercellular Crosstalk Using a 3D Bioprinting Technology

Abstract

Intercellular interaction is known as one of the essential characteristics to recapitulate the intrinsic cellular functions in an engineered system. Among the various shapes of the cell-laden construct, the form of spheroids is suitable for mimicking the minimum unit of the cellular ecosystem in the body. The standard methods of fabricating spheroids deal with the difficulties of producing uniform sized spheroids in a standard high throughput manner and cost considerations. To expedite the fabrication process of size-controlled cell-laden spheroids in a cost-effective way and high throughput production, the present study focused on demonstrating a promising approach to fabricate cell-laden spheroids using three-dimensional (3D) bioprinting technology. By varying parameters such as mixing ratio, nozzle size, and applied pressure, an optimal condition to produce sizecontrolled spheroids were determined. In in vitro, cell-based spheroids demonstrated predominant green fluorescence highlighting the dominated population of live cells and clearly indicates that cells were viable in the biocompatible dECM niche condition. We also will use this printed spheroid platform for co-culturing a multispecies microbial community that shows a cross-feeding metabolism in the human gut in the future.