Self-Organization of Liver Organoids in 3D Liver Extracellular Matrix-Derived Hydrogele

Background: An important advantage of employing extracel-lular matrix (ECM)-derived biomaterials in tissue engineering is the ability to tailor the biochemical and physical microenvi-ronment of the cells. This study aims to assess whether three-dimensional (3D) liver ECM-derived hydrogel (LEMgel) pro-motes physiological functions of self-organized liver organoids generated by human hepatocarcinoma cells together with hu-man mesenchymal and endothelial cells.Materials and Methods: We have fabricated LEMgel with the greatest content of glycosaminoglycans, collagen, laminin and fibronectin. During gelation, complex viscoelasticity modulus of the LEMgel (3 mg/ml) increased from 186.7 to 1570.5 Pa and Tan Delta decreased from 0.27 to 0.18. Scanning electron microscopy (SEM) investigation determined that the LEMgel had a pore size of 382 ± 71 μm.Results: The self-organized organoids in 3D LEMgel (LEMgel-organoid) expressed ALB, CYP3A4, E-cadherin, and ASGPR. The LEMgel-organoid had significant upregulation of transcripts of ALB, CYP3A4, CYP3A7, and TAT as well as downregulation of AFP compared to collagen type I- and scaf-fold-free-organoids or organoids in the LEM emulsion and 2D culture of hepatocarcinoma cells. Generated 3D LEMgel-orga-noids had significantly greater ALB and AAT secretion, urea production, CYP3A4 enzyme activity, and inducibility.Conclusion: In conclusion, liver organoid self-organization within the 3D LEMgel promoted hepatocyte function compared to traditional 2D, 3D, and collagen gel cultures. Our novel 3D LEMgel-organoid could potentially be used in liver tissue engi-neering, drug discovery, or toxicology studies.

کلیدواژه ها:

ECM-Derived Hydrogel ، Human Hepatocarcinoma Cells ، Liver Organoid ، Tissue Engineering