Improving human mesenchymal stem cell-derived hepatic cell energy metabolism by manipulating glucose homeostasis and glucocorticoid signaling

Front Endocrinol (Lausanne). 2023 Jan 13:13:1043543. doi: 10.3389/fendo.2022.1043543. eCollection 2022.


Introduction: The development of reliable hepatic in vitro models may provide insights into disease mechanisms, linking hepatocyte dysmetabolism and related pathologies. However, several of the existing models depend on using high concentrations of hepatocyte differentiation-promoting compounds, namely glucose, insulin, and dexamethasone, which is among the reasons that have hampered their use for modeling metabolism-related diseases. This work focused on modulating glucose homeostasis and glucocorticoid concentration to improve the suitability of a mesenchymal stem-cell (MSC)-derived hepatocyte-like cell (HLC) human model for studying hepatic insulin action and disease modeling.

Methods: We have investigated the role of insulin, glucose and dexamethasone on mitochondrial function, insulin signaling and carbohydrate metabolism, namely AKT phosphorylation, glycogen storage ability, glycolysis and gluconeogenesis, as well as fatty acid oxidation and bile acid metabolism gene expression in HLCs. In addition, we evaluated cell morphological features, albumin and urea production, the presence of hepatic-specific markers, biotransformation ability and mitochondrial function.

Results: Using glucose, insulin and dexamethasone levels close to physiological concentrations improved insulin responsiveness in HLCs, as demonstrated by AKT phosphorylation, upregulation of glycolysis and downregulation of Irs2 and gluconeogenesis and fatty acid oxidation pathways. Ammonia detoxification, EROD and UGT activities and sensitivity to paracetamol cytotoxicity were also enhanced under more physiologically relevant conditions.

Conclusion: HLCs kept under reduced concentrations of glucose, insulin and dexamethasone presented an improved hepatic phenotype and insulin sensitivity demonstrating superior potential as an in vitro platform for modeling energy metabolism-related disorders, namely for the investigation of the insulin signaling pathway.

Keywords: alternative hepatic in vitro models; dexamethasone; glucose; hepatocyte-like cells; insulin; mesenchymal stem cells; metabolism.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Dexamethasone / pharmacology
  • Energy Metabolism
  • Fatty Acids / metabolism
  • Glucocorticoids* / metabolism
  • Glucocorticoids* / pharmacology
  • Glucose / metabolism
  • Hepatocytes / metabolism
  • Homeostasis
  • Humans
  • Insulin / metabolism
  • Mesenchymal Stem Cells* / metabolism
  • Proto-Oncogene Proteins c-akt / metabolism
  • Signal Transduction


  • Glucocorticoids
  • Proto-Oncogene Proteins c-akt
  • Insulin
  • Glucose
  • Dexamethasone
  • Fatty Acids

Grants and funding

The work was financially supported by Fundação para a Ciência e a Tecnologia (FCT) through SFRH/BD/144130/2019 to JRo; PD/BD/114113/2015 and IMM/BI/76-2019 to AF-P; IF/01693/2014 and IMM/CT/27-2020 to VM; PTDC/MED-TOX/29183/2017; UIDB/04138/2020; UIDP/04138/2020 and PTDC/BIACEL/31230/2017. This project has received funding from the European Horizon´s research and innovation programme HORIZON-HLTH-2022-STAYHLTH-02 under agreement No 101095679; from the European Molecular Biology Organization (EMBO-IG#3309); and was also supported by COST actions CA17112, CA20121 and CA20140.