Three-Dimensional Organoids as a Model to Study Nonalcoholic Fatty Liver Disease

Abstract

Despite the rising prevalence of nonalcoholic fatty liver disease (NAFLD), the underlying disease pathophysiology remains unclear. The need is great for an efficient and reliable ‘human’ in vitro model to study NAFLD and the progression to nonalcoholic steatohepatitis (NASH), which will soon become the leading indication for liver transplantation. Here, we review the recent developments in the use of 3D liver organoids as a model to study NAFLD and NASH pathophysiology and possible treatments. Various techniques that are currently used to make liver organoids are discussed, such as the use of induced pluripotent stem cells vs. primary cell lines and human vs. murine cells. Moreover, methods for inducing lipid droplet accumulation and fibrosis to model NAFLD are explored. Finally, the limitations specific to the 3D organoid model for NAFLD/NASH are reviewed, highlighting the need for further development of multilineage models to include hepatic nonparenchymal cells and immune cells. The ultimate goal is to be able to accurately recapitulate the complex liver microenvironment in which NAFLD develops and progresses to NASH.

Experimental models of fatty liver diseases: Status and appraisal

“…Due to the absence of key liver nonparenchymal cell interactions, the results differ significantly from animal models. 117 …”

“…The use of organoids for liver disease research or drug screening has the potential to significantly reduce the number of animal models used for the same purpose. 117 This reduction in the number of cell species, particularly the absence of immune cell involvement, clearly limits the viability of liver organoids for studying complex pathophysiological processes. 126 Thus, the current construction protocol still needs to be improved.…”

The Long Road to Develop Custom-built Livers: Current Status of 3D Liver Bioprinting

“…In recent years, 3D bioprinting of liver organoids has been incorporated into existing organoid technology with promising results. 82 Liver organoids have wide-reaching applications, including cancer 83 and liver disease modeling, 84 drug development, 85 and regenerative medicine. Additionally, bioreactor technology has made impressive technological advancements, 45 and the integration of advanced 3D-printed liver organoid models in conjunction with high-fidelity bioreactors has the potential to revolutionize current liver models (Figure 3).…”

Three-dimensional human bile duct formation from chemically induced human liver progenitor cells

“…Currently, there has been significant improvement in the generation of functional hepatocytes from induced pluripotent stem cells (iPSCs). Several in vitro models of human hepatic disease have been established based on iPSCs ( Olgasi et al, 2020 ; Kim et al, 2022 ; Park et al, 2022 ; Xu et al, 2022 ). Some researchers have also used chemically induced pluripotent stem cells to build various hepatocyte and cholangiocyte organoids ( Si-Tayeb et al, 2010 ; Chen et al, 2018 ; Aizarani et al, 2019 ; Ramli et al, 2020 ; Wang et al, 2020 ; Carberry et al, 2022 ).…”

“…Additionally, the present studies of BDs constructed in vitro usually ignore the relationship between BDs and hepatocytes. Hepatotoxicity studies based on hepatocytes are basically studies of the hepatocyte culture models or cystic hepatic organoids ( Li et al, 2022 ; Park et al, 2022 ). Cystic organoids contain BECs and hepatocytes and exhibit corresponding characteristic cell functions.…”