Three-dimensional (3D) cell culture models, including spheroids and organoids, offer superior physiological relevance compared to 2D systems but remain challenging to scale and standardize. Beyond the complexity of the models themselves, manual handling steps between culture and downstream assays introduce significant variability, limiting reproducibility and complicating protocol transfer across laboratories and platforms.
To address these limitations, we integrated the CellTiter-Glo® 3D Cell Viability Assay into a fully automated, end-to-end workflow on the MO:BOT platform, enabling both 3D model generation and ATP-based viability assessment within a single system. The homogeneous, luminescent CellTiter-Glo® 3D assay (Promega) is inherently compatible with automation and supports robust viability measurements across diverse 3D models, including iPSC-derived cortical, midbrain, and kidney organoids, as well as hepatic spheroids derived from cell lines.
By eliminating manual intervention between culture and assay, the MO:BOT reduces operator-dependent variability and ensures consistent handling conditions across experiments. Automated workflows on MO:BOT consistently yielded higher and less variable CellTiterGlo® 3D signals compared to manual methods, reflecting greater cell viability resulting from standardized cell seeding, medium exchange, and reagent handling throughout the workflow. This integrated approach enables reproducible, transferable, and quality-controlled 3D culture workflows, positioning the MO:BOT as a scalable solution for preclinical research and screening applications.
