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A Superhydrophobic Perforated Microwell Plate for Facile and Robust Spheroid Cultures in Sitting-Drop Format

This paper presents a sitting-drop (SD) strategy for the facile, robust and long-term cell spheroid cultures. Unlike the conventional hanging-drop (HD) method, the SD strategy employs a superhydrophobic perforated microwell plate (SHPMP) for spatially confining arrays of cell suspension droplets to culture 3D spheroids. Benefitting from the combination of upward positioning of cell suspension droplets and non-wettability of microwell surface, the SAHPMP-based SD platform not only maintains the simplicity and high oxygen availability of the HD method but also provides high mechanical stability and easiness of exchanging medium, thus enabling a robust long-term spheroid culture in a high-throughput manner.