A Scalable Platform for Generating Uniform Human-Induced Pluripotent Stem Cell (hiPSC)-Derived Kidney Organoids Using Micropatterning

Background: Organoids hold immense potential for modeling complex diseases, drug discovery, and regenerative medicine. However, current methods for generating organoids suffer from limitations in reproducibility and scalability, hindering their use in high-throughput applications. We present a novel approach to overcome these limitations by developing a standardized micropatterned platform for generating human induced pluripotent stem cell (hiPSC)-derived kidney organoids. Micropatterning has shown promise in improving the development and drug screening capabilities of other organoid types, such as neural organoids. However, its application to hiPSC-derived kidney organoids remains unexplored. Methods: Here, we describe the first-of-its-kind method for generating uniform hiPSC-derived kidney organoids in various micropatterns, including linear, square, and hexagonal shapes, with customizable patterns and sizes. Results: Our micropatterning method successfully generated hiPSC cultures in various geometries, including linear, square, and hexagonal shapes. We achieved precise control over both the pattern and size of the cultured hiPSCs, enabling customization for specific experimental needs. Conclusion: In conclusion, our micropatterning method has established a foundation for the development of standardized hiPSC cultures. The precise control over patterning and size paves the way for tailored experiments and potentially unlocks a new era of consistent and scalable hiPSC-derived organoid generation. We now investigate if micropatterned hiPSCs differentiate into more uniform kidney organoids (size and structure) compared to conventional methods. This analysis, involving comparisons between patterned and non-patterned organoids, will determine if our approach enables large-scale production of consistent kidney organoids for disease modeling, drug screening, and potentially, future kidney regeneration.