Abstract
Bioprinting is a novel technology that may help to overcome limitations associated with two-dimensional (2D) cell cultures and animal experiments, as it allows the production of three-dimensional (3D) tissue models composed of human cells. The present study describes the optimization of a bioink composed of alginate, gelatin and human extracellular matrix (hECM) to print human HepaRG liver cells with a pneumatic extrusion printer. The resulting tissue model was tested for its suitability for the study of transduction by an adeno-associated virus (AAV) vector and infection with human adenovirus 5 (hAdV5). We found supplementation of the basic alginate/gelatin bioink with 0.5 and 1 mg/mL hECM provides desirable properties for the printing process, the stability of the printed constructs, and the viability and metabolic functions of the printed HepaRG cells. The tissue models were efficiently transduced by AAV vectors of serotype 6, which successfully silenced an endogenous target (cyclophilin B) by means of RNA interference. Furthermore, the printed 3D model supported efficient adenoviral replication making it suitable to study virus biology and develop new antiviral compounds. We consider the approach described here paradigmatic for the development of 3D tissue models for studies including viral vectors and infectious viruses.
Highlights
Three-dimensional (3D) bioprinting enables the fabrication of cell-laden biological 3D structures, which can contain multiple cell types as well as different biomaterials within a complex 3D geometry
The present study aims at optimizing conditions using human decellularized extracellular matrix (ECM) as a component of an alginate/gelatin-based bioink, which can be used for extrusion-based bioprinting of a liver model for transduction and infection studies
Our findings demonstrate that supplementation of an alginate/gelatin bioink with 0.5–1 mg/mL human extracellular matrix (hECM) improves cell viability and hepatic metabolic activity (Figures 2C and 3) in a humanized 3D liver model generated by extrusion bioprinting
Summary
Three-dimensional (3D) bioprinting enables the fabrication of cell-laden biological 3D structures, which can contain multiple cell types as well as different biomaterials within a complex 3D geometry. Printed 3D constructs may provide suitable systems to study gene therapeutic strategies as well as tissue-specific infection of different pathogens. As they can be generated with human cells, they may reflect physiological conditions of humans better than animal experiments and thereby help to overcome shortcomings of current models. For a number of human pathogens, no appropriate animal model exists, as there are no natural hosts besides humans. In these cases, repeated passaging may succeed in adapting the pathogens to replicate in animal models; even when successful replication of the virus is achieved, the relevance of these models is often restricted and the pathophysiological manifestation of the infection is limited compared to their human equivalents [1,2,3]. Therapeutic options to treat severe courses of infection are limited, indicating the need to further study AdV biology and develop new antiviral agents
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
