As organoids offer a promising tool to study cell biology and model diseases, organoid technology has rapidly evolved over the last few years. Even though intestinal organoids are one of the most well‐established organoid systems, they currently rely on the embedding into an excess amount of poorly defined, tumor‐derived extracellular matrix. Here, a novel suspension method is suggested to grow mouse intestinal organoids inside thermoformed microwell arrays. This platform promotes the controlled growth of organoids under matrix‐reduced conditions, with Matrigel only used as medium supplement. Hence, this system provides numerous advantages over the previously established methods. Based on the findings, viable and functional mouse intestinal organoids can be preserved for longer periods than in traditional Matrigel domes. Additionally, this microwell‐based technique renders a novel organoid culture system in which the heterogeneity of the organoids is significantly reduced. The method paves the way toward more controlled organoid culture systems that can also be beneficial for further downstream applications, such as automated imaging techniques and micromanipulations, which constitute valuable tools for high‐throughput applications and translational studies.
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