The sexual dimorphism of primary murine hepatocytes changes during cultivation

Question The liver is one of the most sexual dimorphic organs. Cholesterol, fatty acid and drug metabolism are among the signal pathways in the liver that are most affected by sexual dimorphism. This in turn influences the development and course of various liver diseases e.g. non-alcoholic fatty liver disease (NAFLD) and hepatocellular carcinoma (HCC), as well as the general drug treatment in male and female organisms. It is therefore highly important to understand the underlying mechanisms of sexual dimorphism and to take this into account when researching diseases and their treatment. For this reason, we investigated the development of sexual dimorphism in primary hepatocytes during a 96 hour cultivation.

Methods Primary hepatocytes from male and female C57Bl6/N mice were isolated and cultured in collagen coated 6-well plates. Samples were taken at 0, 24, 48, 72 and 96 h. We performed transcriptome, proteome, extracellular metabolome and qPCR analyses to examine the samples.

Results We observed that the gene expression of fatty acid beta oxidation was higher from 0 to 24 h in male compared to female hepatocytes, while it was higher in female hepatocytes from 48 to 96 h. In addition, our results show that the sex specific gene expression of several important cytochrome P450 family members was lost during cultivation e.g. Cyp1a2, Cyp2b23 and Cyp2b9. Furthermore, our data revealed that female hepatocytes consumed more amino acids than male ones. Considering all measured signaling pathways, the sexually dimorphic gene expression pattern of 96 h was most consistent with the initial situation at 0 h.

Conclusions Our results show, how the sex-specific regulation of various central metabolic pathways proceeded during cultivation. Furthermore, the data show that there are large differences in the alteration of sex-specific regulation of drug metabolism during the cultivation of primary hepatocytes, which can have an immense influence on the assessment of pharmacodynamic processes. These results should be taken into account when designing new sex-specific pharmacological liver treatments, which are actually of high interest in the term of personalized medicine.

Publication History

Article published online:
04 January 2021