Multicellular Co-Culture in Three-Dimensional Gelatin Methacryloyl Hydrogels for Liver Tissue Engineering.

Juan Cui,Tao Sun,Qing Shi,Toshio Fukuda,Huaping Wang,Qiang Huang

doi:10.3390/molecules24091762

Juan Cui, Tao Sun + Show 4 more

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https://doi.org/10.3390/molecules24091762

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Abstract

Three-dimensional (3D) tissue models replicating liver architectures and functions are increasingly being needed for regenerative medicine. However, traditional studies are focused on establishing 2D environments for hepatocytes culture since it is challenging to recreate biodegradable 3D tissue-like architecture at a micro scale by using hydrogels. In this paper, we utilized a gelatin methacryloyl (GelMA) hydrogel as a matrix to construct 3D lobule-like microtissues for co-culture of hepatocytes and fibroblasts. GelMA hydrogel with high cytocompatibility and high structural fidelity was determined to fabricate hepatocytes encapsulated micromodules with central radial-type hole by photo-crosslinking through a digital micromirror device (DMD)-based microfluidic channel. The cellular micromodules were assembled through non-contact pick-up strategy relying on local fluid-based micromanipulation. Then the assembled micromodules were coated with fibroblast-laden GelMA, subsequently irradiated by ultraviolet for integration of the 3D lobule-like microtissues encapsulating multiple cell types. With long-term co-culture, the 3D lobule-like microtissues encapsulating hepatocytes and fibroblasts maintained over 90% cell viability. The liver function of albumin secretion was enhanced for the co-cultured 3D microtissues compared to the 3D microtissues encapsulating only hepatocytes. Experimental results demonstrated that 3D lobule-like microtissues fabricated by GelMA hydrogels capable of multicellular co-culture with high cell viability and liver function, which have huge potential for liver tissue engineering and regenerative medicine applications.

Highlights

Construction of alternative liver tissues is urgently required for pharmacological and clinical research [1,2,3]
The gelatin methacryloyl (GelMA) mixed with HepG2 cells was irradiated in a microfluidic channel by UV light that came from the reflection of the digital micromirror device (DMD) illuminated by a mercury lamp
That the GelMA was cross-linked into a radial-pattern micromodule with hepatocytes encapsulation, to mimic liver lobule owing hexagonal-like geometry and radial-like pattern (Figure 1)

Summary

Introduction

Construction of alternative liver tissues is urgently required for pharmacological and clinical research [1,2,3]. Hydrogels are being widely studied to construct cellular microenvironment due to their excellent similarity to the extracellular matrix (ECM) [13,14,15,16] Many hydrogels such as alginate, gelatin, and chitosan have been exploited through various kinds of crosslinking methods for liver tissue engineering [17,18,19]. These studies are focused on cellular sheets constructions with simple structures, which could not Molecules 2019, 24, 1762; doi:10.3390/molecules24091762 www.mdpi.com/journal/molecules

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