Project description:The human body function requires the crosstalk between the central nervous system and its innervated peripheral targets. One such essential crosstalk involves the connections between neural, muscular, and skeletal tissues, which have not yet been modeled with human cells. Here, we describe the generation of three-dimensional, neuromusculoskeletal tri-tissue organoids (hNMSOs) from human pluripotent stem cells. Staining, single-nucleus RNA-sequencing, and spatial transcriptome profiling revealed the co-emergence and spatially confined organization of neural, muscular, and skeletal lineages relevant to human tissues within individual organoids. The neural, muscular, and skeletal regions of hNMSOs exhibited maturation and established functional connections during development, and skeletal support benefited skeletal muscles’ structural and functional development. Modeling with hNMSOs also unveiled the neuromuscular alterations following pathological skeletal degeneration. Together, our study provides an experimental model for future studies of human neuromusculoskeletal crosstalk and disease.
Project description:The human body function requires the crosstalk between the central nervous system and its innervated peripheral targets. One such essential crosstalk involves the connections between neural, muscular, and skeletal tissues, which have not yet been modeled with human cells. Here, we describe the generation of three-dimensional, neuromusculoskeletal tri-tissue organoids (hNMSOs) from human pluripotent stem cells. Staining, single-nucleus RNA-sequencing, and spatial transcriptome profiling revealed the co-emergence and spatially confined organization of neural, muscular, and skeletal lineages relevant to human tissues within individual organoids. The neural, muscular, and skeletal regions of hNMSOs exhibited maturation and established functional connections during development, and skeletal support benefited skeletal muscles’ structural and functional development. Modeling with hNMSOs also unveiled the neuromuscular alterations following pathological skeletal degeneration. Together, our study provides an experimental model for future studies of human neuromusculoskeletal crosstalk and disease.
Project description:The development and function of the human body involves the crosstalk between the central nervous system (CNS) and its innervated peripheral targets. One such essential crosstalk involves the connections between neural, muscular, and skeletal tissues, which have not yet been modeled with human cells. Here, we generate three-dimensional, tri-tissue organoids from human pluripotent stem cells that contain spinal cord, muscular, and skeletal components. The three distinct lineages co-develop in each single organoid and differentiate in a spatially-organized manner. The neural, muscular, and skeletal regions undergo maturation and can establish functional connections during organoid development. Using this system, we find that skeletal support is essential for human skeletal muscles’ structural and functional development. We also show that proinflammatory stimulation in skeletal regions, which resembles the pathological trigger in arthritis-associated conditions, leads to skeletal tissue degeneration, notably, causes neuromuscular structure and function deterioration. We anticipate that our tri-tissue organoids will provide a valuable model for future studies of human neuromusculoskeletal crosstalk and disease.
Project description:Here, we used joint single-nuclei RNA-sequencing (snRNA-seq) and single-nuclei ATAC sequencing (scATAC) to profile freshly isolated crypts from the human fetal intestine and matched intestinal epithelial only organoids (also known as enteroids) derived from these crypts after one passage of in vitro growth. Organoids were grown in the standard 25% LWRN media with either 100 ng/ml of EGF or 1 ng/ml of EREG added. Fresh crypts were not placed in culture but rather immediately frozen for multiomic processing.