Human Hepatic Cell line 5 : In-Vitro Model for Hepatic Immunobiology

Background Hepatocellular carcinoma is a major global health challenge, partly due to the lack of suitable in vitro models that mimic early host–virus interactions. Human Hepatic Cell line 5 (HHL-5), an immortalized hepatocyte cell line, retains key liver functions, lacks tumour markers, binds virus-like particles, and responds to immune stimuli. This study aimed to characterize the genetic and metabolic profile of HHL-5 to evaluate its suitability as a physiologically relevant model for studying viral infection and host immune responses. Method HHL-5 and HepG2 cell lines were analysed for morphology, growth, genetic variants, metabolite profiles, and protein expression. Immunofluorescence and MTS assays assessed cell morphology and growth, while whole exome sequencing and NMR evaluated genetic and metabolic profiles. Protein markers related to proliferation, oxidative stress, and detoxification were examined via Western blot, with significance tested using a T-test. Results Functional analysis of germ line variants in HHL-5 illustrated a highly immunocompetent genomic profile, including antigen processing and presentation, interferon signalling, dendritic cell differentiation, and leukocyte adhesion. Conversely, HepG2 exhibited enrichment in DNA replication pathways. Metabolite analysis in HHL-5 exhibited elevated levels of 1-methylnicotinamide, ADP, and UDP-GalNAc, suggesting enhanced redox function, mitochondrial respiration, and glycosylation—key features of active oxidative metabolism characteristic of primary hepatocytes. In contrast, HepG2 showed increased levels of lactate, glutathione disulfide, creatine, glycerophosphocholine, and branched-chain amino acids, indicative of a glycolytic, redox-adaptive metabolic profile typical of hepatocellular carcinoma. Conclusion HHL-5’s non-cancerous, immunocompetent profile makes it a valuable model for investigating liver disease progression and hepatocarcinogenesis.