Establishing dorsal-ventral patterning in human neural tube organoids with synthetic organizers

Tao Luo¹, Cong Liu², Tao Cheng³, Guo-Qin Zhao², Ying Huang², Jing-Yun Luan⁴, Junyu Guo⁵, Xiang Liu⁴, Yi-Fan Wang⁶, Yang Dong³, Yu Xiao⁷, Enhui He⁸, Rui-Zhen Sun⁹, Xiuyu Chen¹⁰, Jiekai Chen¹⁰, Jun Ma², Sean Megason¹¹, Junfeng Ji¹², Peng-Fei Xu¹³

Affiliations

¹ 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.
² 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.
³ Institute of Genetics, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, China; Women's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310006, China.
⁴ Institute of Genetics, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, China.
⁵ 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.
⁶ Precision Medicine Research Programme, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117599, Singapore.
⁷ College of Computer Science and Technology, Zhejiang University, Hangzhou, China.
⁸ The State Key Laboratory of Brain-Machine Intelligence, Zhejiang University, Hangzhou, China.
⁹ Department of Histology and Embryology, Harbin Medical University, Harbin, Heilongjiang 150081, China.
¹⁰ Center for Biomedical Digital Science, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.
¹¹ Department of Systems Biology, Harvard Medical School, 200 Longwood Avenue, Boston, MA 02115, USA.
¹² 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. Electronic address: jijunfeng@zju.edu.cn.
¹³ 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. Electronic address: pengfei_xu@zju.edu.cn.

PMID: 40373768
DOI: 10.1016/j.stem.2025.04.011

Establishing dorsal-ventral patterning in human neural tube organoids with synthetic organizers

Tao Luo et al. Cell Stem Cell. 2025.

. 2025 Jul 3;32(7):1071-1086.e8.

doi: 10.1016/j.stem.2025.04.011. Epub 2025 May 14.

Authors

Affiliations

¹ 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.
² 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.
³ Institute of Genetics, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, China; Women's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310006, China.
⁴ Institute of Genetics, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310058, China.
⁵ 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.
⁶ Precision Medicine Research Programme, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117599, Singapore.
⁷ College of Computer Science and Technology, Zhejiang University, Hangzhou, China.
⁸ The State Key Laboratory of Brain-Machine Intelligence, Zhejiang University, Hangzhou, China.
⁹ Department of Histology and Embryology, Harbin Medical University, Harbin, Heilongjiang 150081, China.
¹⁰ Center for Biomedical Digital Science, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.
¹¹ Department of Systems Biology, Harvard Medical School, 200 Longwood Avenue, Boston, MA 02115, USA.
¹² 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. Electronic address: jijunfeng@zju.edu.cn.
¹³ 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. Electronic address: pengfei_xu@zju.edu.cn.

PMID: 40373768
DOI: 10.1016/j.stem.2025.04.011

Abstract

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.

Keywords: BMP; SHH; neural tube; organoid; pluripotent stem cells.

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Conflict of interest statement

Declaration of interests The authors declare no competing interests.

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Establishing dorsal-ventral patterning in human neural tube organoids with synthetic organizers

Affiliations

Establishing dorsal-ventral patterning in human neural tube organoids with synthetic organizers

Authors

Affiliations

Abstract

Conflict of interest statement

MeSH terms

LinkOut - more resources

Full Text Sources

Research Materials