Metabolic dynamics of sexual dimorphism in primary mouse hepatocytes in vitro

Background and Aims The liver is one of the most sexually dimorphic organs. Sex-specific differences are evident in metabolism as well as in the development and progression of liver diseases, e. g. non-alcoholic fatty liver disease and hepatocellular carcinoma. Despite these differences, sex of cells is rarely considered in cell culture or development of pharmacological agents. The current project aims to provide a better understanding of molecular mechanisms in sexual dimorphism and their relevance in cell culture experiments.

Method Primary hepatocytes from 12 weeks old female and male C57BL6/N mice were isolated and cultured in collagen-coated plates. Transcriptome, proteome and extracellular metabolome analyses were conducted after 0, 24, 48, 72 and 96 h in vitro cultivation.

Results The sex-specific gene expression of drug metabolism is altered during cultivation. The total amount of sex-specific expressed proteins is greatly reduced with progressing cultivation and partly restored after 96 h. There is significant reduction of female-specific gene expression of Cyp2b13 and Cyp2b9. Sex-dependent differences of several signalling pathways increased, including genes related to serotonin and melatonin degradation. For pathways such as amino acid degradation, beta-oxidation, androgen signalling and hepatic steatosis, the ratios of male and female gene expression were inversed during cultivation.

Conclusion Our results showed large differences in the sex-specific gene and protein expression and the dynamics during cultivation of primary hepatocytes. In consequence of measured signalling pathways, the cultivation time should be adapted to the experimental focus and the dimorphic changes need to be borne in mind for the interpretation of results.

Publikationsverlauf

Artikel online veröffentlicht:
26. Januar 2022