A Sessile Drop Method for Facile and Robust Spheroid Cultures

Abstract In this work, an innovative sessile drop (SD) strategy for facile, robust, and long‐term cell spheroid cultures is proposed. Unlike the conventional hanging drop (HD) method, this strategy employs a superhydrophobic perforated microwell plate (SHPMP) to spatially confine arrays of cell suspension droplets for 3D spheroid cultures. Benefitting from a combination of upward positioning of cell suspension droplets and non‐wettability of the microwell surface, the SD culture platform not only maintains the technical simplicity and high oxygen availability of the HD method, but also provides high mechanical stability and ease of medium exchange, thus enabling a robust long‐term spheroid culture in a high‐throughput manner. In a proof of concept application, the convenience and simplicity of the SD approach is demonstrated by producing 384 uniform spheroids of MHCC97H cell with SHPMP. To validate the utility of this method, it is further demonstrated that spheroids fabricated by this method can be used for drug testing. With convenient liquid handling, low reagent/sample consumption, high droplet stability, adequate oxygen supply, and good in situ monitoring compatibility, the SD platform is beneficial for long‐term cell spheroid culture in a high throughput manner and provides a powerful tool for tissue engineering, regenerative medicine, and drug screening.