Intestinal organoids derived from single cells undergo complex crypt–villus patterning and morphogenesis. However, the nature and coordination of the underlying forces remains poorly characterized. Here, using light-sheet microscopy and large-scale imaging quantification, we demonstrate that crypt formation coincides with a stark reduction in lumen volume. We develop a 3D biophysical model to computationally screen different mechanical scenarios of crypt morphogenesis. Combining this with live-imaging data and multiple mechanical perturbations, we show that actomyosin-driven crypt apical contraction and villus basal tension work synergistically with lumen volume reduction to drive crypt morphogenesis, and demonstrate the existence of a critical point in differential tensions above which crypt morphology becomes robust to volume changes. Finally, we identified a sodium/glucose cotransporter that is specific to differentiated enterocytes that modulates lumen volume reduction through cell swelling in the villus region. Together, our study uncovers the cellular basis of how cell fate modulates osmotic and actomyosin forces to coordinate robust morphogenesis.

来自单细胞的肠类器官经历复杂的隐窝-绒毛模式和形态发生。然而，潜在力量的性质和协调仍不清楚。在这里，使用光片显微镜和大规模成像量化，我们证明隐窝形成与管腔体积的明显减少同时发生。我们开发了一个 3D 生物物理模型来通过计算筛选隐窝形态发生的不同机械场景。将其与实时成像数据和多种机械扰动相结合，我们表明肌动球蛋白驱动的隐窝顶端收缩和绒毛基底张力与管腔体积减少协同作用以驱动隐窝形态发生，并证明了差异张力中存在一个临界点，在该临界点之上，隐窝形态对体积变化变得稳健。最后，我们确定了一种钠/葡萄糖协同转运蛋白，它对分化的肠细胞具有特异性，可通过绒毛区域的细胞肿胀来调节管腔体积的减少。总之，我们的研究揭示了细胞命运如何调节渗透压和肌动球蛋白力以协调稳健的形态发生的细胞基础。