Generation of self-organized neuromusculoskeletal tri-tissue organoids from human pluripotent stem cells
Yin Yao, Wei Zhou, Jinkui Zhu, Ziling Chen, Linlin Jiang, Xuran Zhuang, Jia Chen, Jianfeng Wei, Xiaoxiang Lu, Yantong Liu, Wei Pang, Qinzhi Zhang, Yajing Cao, Zhuoya Li, Yuyan Zhu, Yangfei Xiang
Abstract
The human body function requires crosstalk between different tissues. An essential crosstalk is in the neuromusculoskeletal (NMS) axis involving neural, muscular, and skeletal tissues, which is challenging to model using human cells. Here, we describe the generation of three-dimensional, NMS tri-tissue organoids (hNMSOs) from human pluripotent stem cells through a co-development strategy. Staining, single-nucleus RNA sequencing, and spatial transcriptome profiling revealed the co-emergence and self-organization of neural, muscular, and skeletal lineages within individual organoids, and the neural domains of hNMSOs obtained a ventral-specific identity and produced motor neurons innervating skeletal muscles. The neural, muscular, and skeletal regions of hNMSOs exhibited maturation and established functional connections during development. Notably, structural, functional, and transcriptomic analyses revealed that skeletal support in hNMSOs benefited human muscular development. Modeling with hNMSOs also unveiled the neuromuscular alterations following pathological skeletal degeneration. Together, our study provides an accessible experimental model for future studies of human NMS crosstalk and abnormality. • Human neuromusculoskeletal (NMS) organoids (hNMSOs) are created from hPSCs • Neural, muscular, and skeletal tissues co-develop and self-organize in hNMSOs • Functional connections are established between motor neurons and muscular cells • Skeletal tissue benefits muscular development and NMS disease modeling Modeling the neuromusculoskeletal (NMS) axis with organoids is challenging. Xiang and colleagues developed hPSCs-derived NNMS organoids (hNMSOs) with co-development, self-organization, and functional connection of neural, muscular, and skeletal tissues. hNMSOs revealed skeletal impacts on muscular development and neuromuscular function. These findings provide a model for studying human NMS crosstalk and disease.