Development of a liver tissue organoid

Despite the ability of liver to regenerate from one third its size, little is known about the cellular mechanisms that underpin such an almost limitless potential for regeneration. In vitro culture of cells is often plagued with reductions in cell viability and function, providing limited understanding of regenerative capability outside of in vivo environments. Even in vivo, once sufficient tissue is lost, native regenerative response limits further regeneration capability. Studies using decellularised liver grafts have potential for understanding the mechanisms behind wound healing and repair as well as providing long term evaluation of drug toxicity after seeding due to the native like biological, biochemical and architectural characteristics.
This study aimed to develop a decellularised porcine liver scaffold with native histoarchitecture and biochemical composition, based on the Leeds method, that can support and develop current knowledge of cell seeding methods. Combination of hypo-and hyper-tonic buffers, low concentration SDS (0.1%) and nucleases resulted in effective decellularisation of tissue and maintenance of ECM histoarchitecture. One cycle of SDS (0.1%) showed sufficient cellular and nuclear removal from native porcine liver tissue, creating a biocompatible scaffold that retained overall histoarchitecture. Subsequent investigation of seeding methods identified key areas for development including volume of cell suspension for better cell coverage, analysis of seeding time to increase cell attachment, modification of cell density to alter cell attachment and use of dynamic and static culture conditions to maximise cell penetration. HepaRG and Huh7 cells supported cell attachment for up to 28 days culture, and proliferation for up to 14 and 28 days respectively. Furthermore, functional HepaRG were identified even after 28 day culture. Results indicate the potential benefits of HepaRG and/or Huh7 seeded discs in developing understanding of cell repair, regeneration and wound healing, as well as the dysregulation of matrix remodelling after injury.