Fabrication and Atomic Force Microscopy-Based Mechanical Analysis of Hepatic Organoids

The liver is a vital organ involved in a wide range of processes such as detoxification, protein synthesis, metabolism, and hormone production. Liver diseases, both inherited and viral hepatitis, liver cancer and fatty degeneration are among the leading causes of death in the world. Recent advances in 3D cell culture technology include the use of pluripotent stem cells and adult stem cells that are cultured in vitro to form self-organizing systems. Organoids are self-organizing multicellular structures that reproduce the structure and function of organs and can be used to model the development, maintenance and repair of organs ex vivo. That is why the search for new methods for the formation of hepatic organoids seen as an urgent task. For the fabrication of 3D organoids we isolated hepatic duct cells from rat liver. The obtained cellular structures were analyzed using atomic force microscopy to estimate their mechanical properties and demonstrated increasing of Young Modulus in comparison with normal liver sections. Morphometric evaluation showed that extracellular matrix fibers occupy up to 60% of the organoid, while cell agglomerations up to 40% of it. We observed the spontaneously formed fibrotic liver tissue-like constructs within 21 days. So, obtained hepatic organoids characterized by a tissue-like structure with a predominance of extracellular matrix fibers in its composition similar to liver tissue affected by fibrosis.