Ruider Iris, Pastucha Anna, Raich Marion K, Xu Wentao, Liu Yan, Reichert Maximilian, Weitz David, Bausch Andreas R
Department of Bioscience, TUM School of Natural Sciences, Heinz Nixdorf Chair in Biophysical Engineering of Living Matter, Technical University of Munich, 85748 Garching, Germany.
Center for Functional Protein Assemblies (CPA), Technical University of Munich, 85748 Garching, Germany.
Lab Chip. 2025 Aug 20. doi: 10.1039/d5lc00287g.
Droplet-based organoid culture offers several advantages over conventional bulk organoid culture, such as improved yield, reproducibility, and throughput. However, organoids grown in droplets typically display only a spherical geometry and lack the intricate structural complexity found in native tissue. By incorporating singularised pancreatic ductal adenocarcinoma cells into collagen droplets, we achieve the growth of branched structures, indicating a more complex interaction with the surrounding hydrogel. A comparison of organoid growth in droplets of different diameters showed that while geometrical confinement improves organoid homogeneity, it also impairs the formation of more complex organoid morphologies. Thus, only in 750 μm diameter collagen droplets did we achieve the consistent growth of highly branched structures with a morphology closely resembling the structural complexity achieved in traditional bulk organoid culture. Moreover, our analysis of organoid morphology and transcriptomic data suggests an accelerated maturation of organoids cultured in collagen droplets, highlighting a shift in developmental timing compared to traditional systems.
基于微滴的类器官培养相对于传统的批量类器官培养具有多个优势，例如产量提高、可重复性增强和通量增加。然而，在微滴中生长的类器官通常仅呈现球形几何形状，缺乏天然组织中存在的复杂结构复杂性。通过将单个胰腺导管腺癌细胞整合到胶原微滴中，我们实现了分支结构的生长，这表明与周围水凝胶存在更复杂的相互作用。对不同直径微滴中类器官生长的比较表明，虽然几何限制提高了类器官的同质性，但也会损害更复杂类器官形态的形成。因此，只有在直径750μm的胶原微滴中，我们才实现了高度分支结构的一致生长，其形态与传统批量类器官培养中实现的结构复杂性非常相似。此外，我们对类器官形态和转录组数据的分析表明，在胶原微滴中培养的类器官成熟加速，突出了与传统系统相比发育时间的转变。
