Luo Tao, Liu Cong, Cheng Tao, Zhao Guo-Qin, Huang Ying, Luan Jing-Yun, Guo Junyu, Liu Xiang, Wang Yi-Fan, Dong Yang, Xiao Yu, He Enhui, Sun Rui-Zhen, Chen Xiuyu, Chen Jiekai, Ma Jun, Megason Sean, Ji Junfeng, Xu Peng-Fei
Center of Stem Cell and Regenerative Medicine, and Bone Marrow Transplantation Center of the First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, China; Nanhu Brain-Computer Interface Institute, Hangzhou, Zhejiang 311100, China.
Center for Genetic Medicine, the Fourth Affiliated Hospital of School of Medicine, and International School of Medicine, Zhejiang University, Yiwu 322000, China; Institute of Genetics, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, China.
Cell Stem Cell. 2025 Jul 3;32(7):1071-1086.e8. doi: 10.1016/j.stem.2025.04.011. Epub 2025 May 14.
Precise dorsal-ventral (D-V) patterning of the neural tube (NT) is essential for the development and function of the central nervous system. However, existing models for studying NT D-V patterning and related human diseases remain inadequate. Here, we present organizers derived from pluripotent stem cell aggregate fusion ("ORDER"), a method that establishes opposing BMP and SHH gradients within neural ectodermal cell aggregates. Using this approach, we generated NT organoids with ordered D-V patterning from both zebrafish and human pluripotent stem cells (hPSCs). Single-cell transcriptomic analysis revealed that the synthetic human NT organoids (hNTOs) closely resemble the human embryonic spinal cord at Carnegie stage 12 (CS12) and exhibit greater similarity to human NT than to mouse models. Furthermore, using the hNTO model, we demonstrated the critical role of WNT signaling in regulating intermediate progenitors, modeled TCTN2-related D-V patterning defects, and identified a rescue strategy.
神经管（NT）精确的背腹（D-V）模式对于中枢神经系统的发育和功能至关重要。然而，现有的用于研究NT D-V模式及相关人类疾病的模型仍存在不足。在此，我们展示了源自多能干细胞聚集体融合的组织者（“ORDER”），这是一种在神经外胚层细胞聚集体中建立相反的骨形态发生蛋白（BMP）和音猬因子（SHH）梯度的方法。利用这种方法，我们从斑马鱼和人类多能干细胞（hPSC）中生成了具有有序D-V模式的NT类器官。单细胞转录组分析表明，合成的人类NT类器官（hNTO）与卡内基阶段12（CS12）的人类胚胎脊髓非常相似，并且与人类NT的相似性高于小鼠模型。此外，使用hNTO模型，我们证明了WNT信号在调节中间祖细胞中的关键作用，模拟了与TCTN2相关的D-V模式缺陷，并确定了一种挽救策略。
