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.
Highlights
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
The g-force, which refers to the relative centrifuge force (RCF), is expressed as REF = 1.118 × r ×2 and was adopted to estimate the g-force value employed in previously published protocols using either an orbital shaker or a magnetic stirrer[25,26,27,28,29,30]
The resultant 3D-human liver scaffolds are characterized by the preservation of essential biochemical, physical and topographical properties and in addition provide an optimal platform for tissue bio-engineering employing different types of human liver cells
Summary
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. 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. Biomaterials should be able to reproduce the essential characteristics of the physiological extracellular scaffold including tissue-specific ECM protein composition, 3D-microarchitecture, stiffness and pro-angiogenic properties in order to support cellular growth and maintain cell phenotypes. Decellularization is a process that removes cellular and immunogenic materials from tissues and organs while maintaining the mechanical and bioactive properties of the tissue[16] This provides the optimal micro-environment for the repopulation with organ-specific cell types leading to the development of engineered tissues and organs. Major advances in the decellularization-recellularization technology have been achieved for the development of whole engineered organs due to their attractive application in the area of whole organ transplantation[1, 2, 17,18,19,20,21]
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.
