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Hepatocellular carcinoma (HCC) possesses distinctive tumorigenic mechanisms, a characteristic tumor microenvironment, robust angiogenic capacity and prominent chemoresistance. These biological traits endow HCC with high tumor heterogeneity, which greatly complicates basic mechanistic research and clinical intervention. At present, there is a shortage of micro-morphological and molecular biomarkers for early clinical screening. Early detection of HCC mainly relies on imaging examinations, which are limited by delayed diagnosis and insufficient sensitivity. Compared with conventional animal experimental models, organoids can steadily retain human-specific biological and histological features and accurately recapitulate the genomic, epigenomic and microenvironmental landscapes of primary tumors, serving as an ideal in vitro research system for screening early diagnostic markers and prognostic factors for tumors. By reconstructing the extracellular matrix microenvironment required for liver development and regeneration, supplemented with key cytokines including hepatocyte growth factor (HGF), epidermal growth factor (EGF), fibroblast growth factor (FGF) and R‑Spondin 1, relevant studies have effectively overcome the technical bottleneck of poor long-term expansion of adult stem cell-derived organoids and successfully established a culture system for normal liver organoids capable of stable long-term passaging. At present, HCC organoids have been widely adopted to dissect the mechanisms underlying malignant tumor progression and provide novel technical support for research on individualized liver cancer therapy. Furthermore, this model can faithfully recapitulate the early evolutionary stages of tumorigenesis, facilitating the discovery of novel molecular markers for early diagnosis as well as the screening and validation of key regulatory genes governing HCC progression, invasion and malignant transformation.