Enormous amounts of essential intracellular events are crowdedly packed inside picoliter-sized cellular space. However, the significance of the physical properties of cells remains underappreciated because of a lack of evidence of how they affect cellular functionalities. Here, we show that volumetric compression regulates the growth of intestinal organoids by modifying intracellular crowding and elevating Wnt/β-catenin signaling. Intracellular crowding varies upon stimulation by different types of extracellular physical/mechanical cues and leads to significant enhancement of Wnt/β-catenin signaling by stabilizing the LRP6 signalosome. By enhancing intracellular crowding using osmotic and mechanical compression, we show that expansion of intestinal organoids was facilitated through elevated Wnt/β-catenin signaling and greater intestinal stem cell (ISC) self-renewal. Our results provide an entry point for understanding how intracellular crowdedness functions as a physical regulator linking extracellular physical cues with intracellular signaling and potentially facilitate the design of engineering approaches for expansion of stem cells and organoids.

大量重要的细胞内事件拥挤在皮升大小的细胞空间内。然而，由于缺乏关于它们如何影响细胞功能的证据，细胞物理特性的重要性仍未得到充分认识。在这里，我们展示了体积压缩通过改变细胞内拥挤和提升 Wnt/β-catenin 信号来调节肠道类器官的生长。细胞内拥挤因不同类型的细胞外物理/机械信号的刺激而变化，并通过稳定 LRP6 信号体导致 Wnt/β-连环蛋白信号的显着增强。通过使用渗透和机械压缩增强细胞内拥挤，我们表明，通过升高的 Wnt/β-catenin 信号传导和更大的肠道干细胞 (ISC) 自我更新促进了肠道类器官的扩张。我们的研究结果为理解细胞内拥挤如何作为连接细胞外物理信号与细胞内信号传导的物理调节器发挥作用提供了一个切入点，并可能有助于设计用于扩增干细胞和类器官的工程方法。