Biofabrication of perfusable liver constructs

Abstract

Event Abstract Back to Event Biofabrication of perfusable liver constructs Pedro F. Costa1, Monique Schuddeboom2, Ferry Melchels1, Jos Malda1, 3 and Bart Spee2 1 University Medical Center Utrecht, Department of Orthopaedics, Netherlands 2 Utrecht University, Clinical Sciences of Companion Animals, Netherlands 3 Utrecht University, Equine Sciences, Netherlands Introduction: Current in vitro systems do not allow an accurate prediction of drug-induced liver injury (DILI). This is one of the main reasons why DILI still occurs in clinical phases of drug development or even post-marketing, which makes it a serious health concern. Our approach is to develop a biofabricated 3D liver construct that can be cultured in a custom designed bioreactor system. As such these perfusable constructs will better mimic the liver compared to a 2D hepatocyte culture. To achieve this, biofabrication techniques were employed with liver cell lines and gelMA hydrogel in a 3D bioprinter. Materials and Methods: As a source of hepatocytes, adult stem cells of the liver, termed liver organoids, or an hepatic cell line (Huh7) are used in the liver constructs. The creation of aggregates of liver organoids and multipotent stromal cells derived from liver (LMSCs) are believed to increase the performance of the liver constructs as determined by hepatic function assays and gene-expression profiling. Bioreactors were custom designed and produced by stereolithography technology. Liver cells were printed in 5% w/v gelMA hydrogel and subsequently cross-linked by UV-A irradiation. The simultaneous deposition of pluronic F127 as a sacrificial support material, allows the formation of a porous structure. After printing, liver constructs are followed over time with cell viability assays including Alamar Blue and life-dead stainings. Cellular damage after treatment with toxic compounds is determined with an ATP assay. Results and discussion: Cell aggregates remained viable for at least four days after 3D printing. The combination of liver organoids and LMSC aggregates in gelMA increased the albumin expression compared to single cell aggregates. However, gene expression profiling of the single cell aggregates with only organoids showed a higher expression of early and late hepatic markers compared to the combined aggregates with organoids and LMSCs. No increased hepatic function of organoid and LMSC containing aggregates was observed with cytochrome P450 assays. As a control, acute toxicity of the liver constructs was determined with a four-fold increase in ATP levels after Triton X-100 treatment at 2-3 days post printing. Placement of the porous liver construct in the custom designed bioreactor allowed the perfusion of the liver construct. Conclusion: These results indicate that perfusable liver bioreactor systems have the potential to better predict DILI. Keywords: Tissue Engineering, biomaterial, Bioprinting, Drug testing Conference: 10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016. Presentation Type: Poster Topic: Biomaterials in constructing tissue substitutes Citation: Costa PF, Schuddeboom M, Melchels F, Malda J and Spee B (2016). Biofabrication of perfusable liver constructs. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.02856 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 27 Mar 2016; Published Online: 30 Mar 2016. Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Pedro F Costa Monique Schuddeboom Ferry Melchels Jos Malda Bart Spee Google Pedro F Costa Monique Schuddeboom Ferry Melchels Jos Malda Bart Spee Google Scholar Pedro F Costa Monique Schuddeboom Ferry Melchels Jos Malda Bart Spee PubMed Pedro F Costa Monique Schuddeboom Ferry Melchels Jos Malda Bart Spee Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.