Abstract

The generation of human pluripotent stem cell (hPSC)-derived kidney organoids has provided an unprecedented platform for modeling human development and disease. This webinar will discuss several key advances in kidney organoid generation that enhance organoid differentiation, improve their resemblance to native kidney tissue, and enable their reproducible production at scale. These advances include bioengineering strategies that promote organoid vascularization by modulating the cellular microenvironment through the use of hydrogels. In addition, a reproducible and cost-effective free-floating culture protocol for generating diverse renal cell types will be presented in detail. This protocol incorporates defined culture conditions that promote the differentiation of kidney organoids into segmented nephron structures containing multiple renal cell types. These outcomes have been comprehensively characterized using a range of analytical techniques, including single-cell RNA sequencing, confocal microscopy with quantitative image analysis, and metabolic profiling.

Highlights:

• Step-by-step generation of human iPSC-derived kidney organoids in static and suspension culture.

• Establishment of natural hydrogels derived from decellularized extracellular matrix to promote in vitro vascularization.

• Functional and structural validation using single-cell RNA sequencing, confocal image analysis, metabolic assays, and CRISPR-Cas9 engineering.

• Applications in studying kidney development, modeling kidney diseases, and ex vivo cell-based therapies in clinical trials.