Transcriptomic Characterisation of Mouse Embryonic Stem Cell-Derived Hepatocyte-Like Cells Generated Using an Advanced Hepatic-Inducing Cocktail (A-HIC)

Bulk RNA sequencing was performed on four independently generated A-HIC-derived HLC samples. Transcriptomic profiles were benchmarked against publicly available hepatic reference datasets to assess their relationship to mouse hepatocyte development and to the AML12 mouse hepatocyte cell line. Weighted gene co-expression network analysis (WGCNA) was used to identify co-expression modules and compare module-level expression patterns with purified mouse hepatocytes representing developmental stages E11.5, E15.5, P1, P10, P30 and week 8.

Developmental-stage associations were further evaluated using Spearman correlation analysis and hepatic marker gene expression. A-HIC-derived HLCs displayed transcriptomic characteristics spanning foetal-to-postnatal hepatocyte development, with the strongest overall correspondence extending from late foetal to early postnatal stages. Of the genes analysed in the developmental comparison, 75.3% of A-HIC genes could be associated with a developmental stage, compared with 45.0% for AML12 cells.

Developmental-stage mapping and hepatic marker expression therefore indicated broader and more advanced developmental alignment of A-HIC-derived HLCs than AML12 cells. Together, the functional and transcriptomic analyses support the hepatic identity and functional maturation of A-HIC-derived HLCs. These RNA-seq data provide a resource for characterising mESC-derived hepatocyte-like cells and assessing their developmental maturity relative to established mouse hepatic reference models.