Reconfigurable Hanging Drop Microarray Platform for On‐Demand Preparation and Analysis of Spheroid Array

Abstract

In response to the increasing demand for spheroid‐based cancer research, the importance of developing integrated platforms that can simultaneously facilitate high‐throughput spheroid production and multiplexed analysis has been emphasized. In addition, the understanding of how the size and cellular composition of tumors directly influence their internal structures and functionalities underlines the critical need to produce spheroids of diverse sizes and compositions on a large scale. To address this rising demand, we present a configurable and linkable in vitro 3D cell culture kit for spheroids, termed CLiCK‐Spheroid. This platform consists of three primary components: a hanging drop microarray (HDMA), concave pillar microarray (CPMA), and gradient blocks. The HDMA alone produces a homogeneous spheroid array, while its combination with the gradient block enables one‐step generation of a size‐gradient spheroid array. Using the size‐gradient spheroid arrays, the occurrence of necrotic cores based on spheroid size is demonstrated. Additionally, spheroids in a single batch can be conveniently compartmentalized and regrouped using CPMA, enhancing the versatility of spheroid arrays and enabling multiplexed drug treatments. By combining the different assembly methods, we have achieved a high‐throughput production of cell composition‐gradient spheroid arrays, with noticeable variations in morphology and vascularization based on cell compositions.This article is protected by copyright. All rights reserved