Introduction:
Kidney injury (KI) is a leading public health issue associated with increased morbidity and mortality. Drug-induced nephrotoxicity (DIN) is a common cause of KI that affects kidney structure and function and presents a major clinical problem. The high incidence of DIN is mainly due to the lack of reliable platforms for nephrotoxicity screening during pre-clinical drug development and clinical trials. Recent advances of pluripotent stem cell-based 2D/3D differentiation approaches and the development of 3D kidney in vitro/ex vivo models have opened up the possibility of novel drug screening platforms. However, it remains to be elucidated whether the stem cell types generated in 2D or 3D have the equivalent renal differentiation potential.
Methods:
In this study, we generated a Rosa26-E2-Crimson/T-GFP dual-labelled mouse embryonic stem cell line and differentiated the cells under 2D monolayer and 3D typical embryoid body culture conditions, respectively. E2-Crimson-labelled nascent mesodermal cells (T-GFP+-expressing populations) were isolated and analysed for the expression of renal-specific genes. Their nephrogenic potential was also investigated by incorporating them within an ex vivo 3D self-assembly kidney organoid model.
Results:
Key nephrogenic genes expression patterns and nephron features including glomeruli and proximal tubules were confirmed. It was also revealed that the majority of T-GFP+ mesodermal cells arising from 3D culture condition displayed a vascular progenitor-like property and eventually differentiated into more mature endothelial cells forming interconnected networks in the organoid model. In contrast, the nascent mesodermal cells derived from 2D culture system appeared to develop into the renal stroma.
Conclusions:
This study showed distinct nephrogenic potential between nascent mesodermal cells arising in 3D and 2D differentiation culture. The self-assembly kidney organoid model provides new insights into the development of renal microphysiological platforms that could more reliably mimic the in vivo environment in nephrotoxicity screening.
I have no potential conflict of interest to disclose.
I did not use generative AI and AI-assisted technologies in the writing process.