Rapid production of human liver scaffolds for functional tissue engineering by high shear stress oscillation-decellularization

Mazza, Giuseppe; Al-Akkad, Walid; Telese, Andrea; Longato, Lisa; Urbani, Luca; Robinson, Benjamin; Hall, Andrew; Kong, Kenny; Frenguelli, Luca; Marrone, Giusi; Willacy, Oliver; Shaeri, Mohsen; Burns, Alan; Malago, Massimo; Gilbertson, Janet; Rendell, Nigel; Moore, Kevin; Hughes, David; Notingher, Ioan; Jell, Gavin; Del Rio Hernandez, Armando; De Coppi, Paolo; Rombouts, Krista; Pinzani, Massimo

Abstract

The development of human liver scaffolds retaining their 3-dimensional structure and extra-cellular matrix (ECM) composition is essential for the advancement of liver tissue engineering. We report the design and validation of a new methodology for the rapid and accurate production of human acellular liver tissue cubes (ALTCs) using normal liver tissue unsuitable for transplantation. The application of high shear stress is a key methodological determinant accelerating the process of tissue decellularization while maintaining ECM protein composition, 3D-architecture and physico-chemical properties of the native tissue. ALTCs were engineered with human parenchymal and non-parenchymal liver cell lines (HepG2 and LX2 cells, respectively), human umbilical vein endothelial cells (HUVEC), as well as primary human hepatocytes and hepatic stellate cells. Both parenchymal and non-parenchymal liver cells grown in ALTCs exhibited markedly different gene expression when compared to standard 2D cell cultures. Remarkably, HUVEC cells naturally migrated in the ECM scaffold and spontaneously repopulated the lining of decellularized vessels. The metabolic function and protein synthesis of engineered liver scaffolds with human primary hepatocytes reseeded under dynamic conditions were maintained. These results provide a solid basis for the establishment of effective protocols aimed at recreating human liver tissue in vitro.

Citation

Mazza, G., Al-Akkad, W., Telese, A., Longato, L., Urbani, L., Robinson, B., Hall, A., Kong, K., Frenguelli, L., Marrone, G., Willacy, O., Shaeri, M., Burns, A., Malago, M., Gilbertson, J., Rendell, N., Moore, K., Hughes, D., Notingher, I., Jell, G., …Pinzani, M. (in press). Rapid production of human liver scaffolds for functional tissue engineering by high shear stress oscillation-decellularization. Scientific Reports, 7, Article 5534. https://doi.org/10.1038/s41598-017-05134-1

• Journal Article Type: Article
• Acceptance Date: Jun 9, 2017
• Online Publication Date: Jul 17, 2017
• Deposit Date: Oct 10, 2017
• Publicly Available Date: Oct 10, 2017
• Journal: Scientific Reports
• Electronic ISSN: 2045-2322
• Publisher: Nature Publishing Group
• Peer Reviewed: Peer Reviewed
• Volume: 7
• Article Number: 5534
• DOI: https://doi.org/10.1038/s41598-017-05134-1
• Keywords: Biomaterials, Tissue engineering
• Public URL: https://nottingham-repository.worktribe.com/output/872868
• Publisher URL: https://www.nature.com/articles/s41598-017-05134-1
• Contract Date: Oct 10, 2017