Establishment of a 3D hyperuricemia model based on cultured human liver organoids

Hyperuricemia (HUA) is a metabolic disease caused by abnormal purine metabolism, the prevalence of which has increased worldwide. Here, a 3D organoid culture system for mimicking HUA in vitro was established using cultured human liver organoids. Liver organoids can be generated from single hepatocytes and passaged for several months, retaining key morphological features, functional purine metabolism and global gene expression profile. Furthermore, organoids can be differentiated into hepatocytes with high expression of maturation markers including HNF4a, E-cadherin, and ALB. Importantly, organoids can produce high level of uric acid after xanthine induction which is the substrate of xanthine oxidase. Furthermore, the preclinical application potential of this organoid model was verified by measuring the anti-hyperuricemic effect of the widely used allopurinol, as well as the reported bioactive substance puerarin. The results demonstrate that this novel organoid model could be used for high-throughput screening of both chemical and food-derived compounds with antihyperuricemic bioactivity. Overall design: Primary Hepatocytes were isolated from paracarcinomatous liver tissues by two-step collagenase digestion, and suspended the cells in Matrigel. The mRNA profile of human liver organoids after 5 days of differentiation and amplification. Total RNA was extracted from 3 liver-organoids from different passages in expansion medium (EM) and differentiation medium (DM) (P4, P6, P8) using TRIzol reagent. RNA quality and quantity using was analyzed by NanoDrop spectrophotometer (BioRad, USA) and a Bioanalyzer 2100 system (Agilent, USA). Transcriptional sequencing library was constructed by TruSeq™ RNA Sample Preparation Kit (Illumina, USA). The libraries were then sequenced on an Illumina Hiseq platform. Sequencing data analysis was performed using the DESeq2 package (CIT) in the R software environment. HEM represents organoids cultured in EM, and HDM represents organoids cultured in DM.