Self-buoyant cell-spheroid culture programming multi-spheroid assembly

Abstract

Tempo-spatially controlled culture and assembly of cellular spheroids are fundamental in engineering tissue-like assembloids for scalable application, yet remain a technical challenge. Here, we show self-buoyant culture of cell spheroids and programmable assembly of multi-spheroid by microbubbles in a high-throughput, external field-free and miniaturized format. This approach allows one-step engineering of biocompatible cell-adhesive buoyancy interface that lifts the self-adaptive levitation growth of reliable one-drop-one-spheroid in an array without requiring any external fields. Using this self-floating spheroidal microarray, we find that a facile droplet ‘kiss’ facilitates the buoyancy-driven ultrafast transfer (∼1 s) of spheroids between top-down adjacent droplets, thus achieving rapid spheroid relocation, media exchange, and drug administration in parallel. Such a self-buoyant approach allows programmable horizontal/vertical bioassembly and enhanced fusion of homo/heterogeneous multi-spheroids into different “buoyantoid” patterns via sequential flash transfer along droplet array. This self-powered design provides a promising tool to enable mass production and smart manipulation of encoded assembloids for precision medicine, tissue engineering and high-throughput drug screening.