Department of Engineering University of Cambridge Cambridge CB2 1PZ UK.
Wellcome - MRC Cambridge Stem Cell Institute University of Cambridge Cambridge CB2 0AW UK.
Adv Sci (Weinh). 2021 Feb 8;8(9):2003332. doi: 10.1002/advs.202003332. eCollection 2021 May.
Epithelial, stem-cell derived organoids are ideal building blocks for tissue engineering, however, scalable and shape-controlled bio-assembly of epithelial organoids into larger and anatomical structures is yet to be achieved. Here, a robust organoid engineering approach, Multi-Organoid Patterning and Fusion (MOrPF), is presented to assemble individual airway organoids of different sizes into upscaled, scaffold-free airway tubes with predefined shapes. Multi-Organoid Aggregates (MOAs) undergo accelerated fusion in a matrix-depleted, free-floating environment, possess a continuous lumen, and maintain prescribed shapes without an exogenous scaffold interface. MOAs in the floating culture exhibit a well-defined three-stage process of inter-organoid surface integration, luminal material clearance, and lumina connection. The observed shape stability of patterned MOAs is confirmed by theoretical modelling based on organoid morphology and the physical forces involved in organoid fusion. Immunofluorescent characterization shows that fused MOA tubes possess an unstratified epithelium consisting mainly of tracheal basal stem cells. By generating large, shape-controllable organ tubes, MOrPF enables upscaled organoid engineering towards integrated organoid devices and structurally complex organ tubes.
上皮细胞干细胞衍生类器官是组织工程的理想构建模块，然而，将上皮细胞类器官大规模地、可控地组装成更大和更具解剖结构的类器官仍然尚未实现。在这里，提出了一种稳健的类器官工程方法，即多类器官图案化和融合（Multi-Organoid Patterning and Fusion，MOrPF），用于将不同大小的气道类器官组装成具有预定形状的无支架、规模化的气道管。多类器官聚集体（Multi-Organoid Aggregates，MOAs）在基质耗尽的、自由漂浮的环境中加速融合，具有连续的腔，并在没有外源性支架界面的情况下保持规定的形状。在悬浮培养中，MOAs 表现出明确的三个阶段的过程，包括类器官表面整合、腔材料清除和腔连接。基于类器官形态和类器官融合涉及的物理力的理论模型证实了图案化 MOAs 的形状稳定性。免疫荧光表征表明，融合的 MOA 管具有无分层的上皮组织，主要由气管基底干细胞组成。通过生成大尺寸、形状可控的器官管，MOrPF 能够实现规模化的类器官工程，以构建集成的类器官装置和具有复杂结构的器官管。
