A Robotic Micromanipulation System for Homogeneous Organoid Culture

Organoids are cell clusters cultured in vitro that maintain the structure and function of the donor organs. They have found important applications in biomedicine, such as drug screening and personalized therapy. However, conventional organoid culture methods lack control of physical properties like size and distribution, leading to increased heterogeneity and very low batch-to-batch reproducibility, which significantly limits their widespread use. Controlling these properties at the microscale is challenging, particularly for fragile fragments, which are the main source for culturing organoids. To address this issue, we present a robotic micromanipulation system that allows operators to select fragments of particular sizes and automatically transfer them into a customized in-situ organoid chip (IOC) for culture. The chip was designed with microwell arrays to uniform the culture environment and facilitate imaging analysis. The transfer of fragments is modeled based on computational fluid dynamics (CFD) and is enabled by designing a robust model predictive control (RMPC) framework. Simulation and experiment results demonstrated the effectiveness of the model and controller. In colorectal cancer organoid culture experiments, our system significantly improved the morphological homogeneity of organoids. Note to Practitioners-Organoids have been demonstrated to be one of the most promising in vitro models. Lacking control of its size and distribution results in significant heterogeneity and low batch-to-batch reproducibility, which limits its wide uses. Here, we report a robotic micromanipulation system that allows operators to select fragments of particular sizes and morphologies and automatically transfer them into a customized organoid chip for culture. The results of colorectal cancer organoids culture experiments verified the effectiveness of our system in reducing the morphological heterogeneity among organoids.