HepaRG细胞自组织形成肝类器官的动态转录物组研究OA

Objective To investigate the time-dependent transcriptomic alterations during liver organoid formation from HepaRG cells under three-dimensional culture from 0 to 120 h,and to characterize molecular features associ-ated with the programmatic transition from proliferation-related processes to hepatic functional differentiation.Methods HepaRG cells were mixed with Matrigel(1∶1,v/v)and seeded into ultra-low-attachment 96-well plates.Samples collected at 0 h,14 h,24 h,72 h,and 120 h were subjected to RNA sequencing(RNA-seq),followed by principal component analysis(PCA)/correlation assessment,DESeq2-based differential expression,time-course clustering with enrichment analysis,and protein-protein interaction(PPI)network analysis.Results HepaRG cells self-assembled into spheroid organoids under three-dimensional culture,accompanied by increased albumin(ALB)expression.The number of differentially expressed genes(DEGs)peaked at 72 h and 120 h.Six tem-poral clusters revealed an early-to-late transition from proliferation/cell-cycle programs to hepatic functional programs(lipid metabolism,lipoprotein remodeling,and the complement pathway).Conclusions HepaRG organoid formation involves dynamic transcriptomic remodeling.The 72-120 h interval constitutes a critical stage with enhanced hepatic function-related programs,offering transcriptomic support for maturation evaluation and mechanistic studies.