Stem Cell Reports. 2024 Sep 10;19(9):1289-1303. doi: 10.1016/j.stemcr.2024.07.010
In vitro differentiation of mouse pluripotent stem cells into corticosteroid-producing adrenocortical cells
Ioannis Oikonomakos1, Melina Tedesco2, Fariba Jian Motamedi2, Mirko Peitzsch3, Serge Nef4, Stefan R Bornstein5, Andreas Schedl6, Charlotte Steenblock7, Yasmine Neirijnck2
Affiliations
1Université Côte d’Azur, Inserm, CNRS, Institut de Biologie Valrose, 06108 Nice, France; Department of Internal Medicine III, University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany.
2Université Côte d’Azur, Inserm, CNRS, Institut de Biologie Valrose, 06108 Nice, France.
3Institute of Clinical Chemistry and Laboratory Medicine, University Hospital Carl Gustav Carus, Medical Faculty Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany.
4Department of Genetic Medicine and Development, University of Geneva, 1211 Geneva, Switzerland.
5Department of Internal Medicine III, University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany.
6Université Côte d’Azur, Inserm, CNRS, Institut de Biologie Valrose, 06108 Nice, France.
7Department of Internal Medicine III, University Hospital Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany.
Abstract
Directed differentiation of pluripotent stem cells into specialized cell types represents an invaluable tool for a wide range of applications. Here, we have exploited single-cell transcriptomic data to develop a stepwise in vitro differentiation system from mouse embryonic stem cells into adrenocortical cells. We show that during development, the adrenal primordium is embedded in an extracellular matrix containing tenascin and fibronectin. Culturing cells on fibronectin during differentiation increased the expression of the steroidogenic marker NR5A1. Furthermore, 3D cultures in the presence of protein kinase A (PKA)-pathway activators led to the formation of aggregates composed of different cell types expressing adrenal progenitor or steroidogenic markers, including the adrenocortical-specific enzyme CYP21A1. Importantly, in-vitro-differentiated cells responded to adrenocorticotropic hormone (ACTH) and angiotensin II with the production of glucocorticoids and mineralocorticoids, respectively, thus confirming the specificity of differentiation toward the adrenal lineage.
DOI: 10.1016/j.stemcr.2024.07.010