The increasing shift toward personalized medicine, coupled with regulatory changes such as the FDA Modernization Act 2.0, is accelerating the transition to 3D cell culture as an alternative to animal models. Automation is essential in supporting this shift by enabling reproducible, scalable, and low-touch workfl ows. The Cell Care STAR now expands its capabilities to include the automated generation and maintenance of 3D cell structures. This Application Note demonstrates how the “Seeding from trough 3D” and “Refresh 3D” work steps enable robust spheroid formation and culture, using the Akura™ 96 Spheroid Microplate (InSphero) as a model system.