Spatially Defined Single-Cell Transcriptional Profiling Characterizes Diverse Chondrocyte Subtypes and Nucleus Pulposus Progenitors in Human Intervertebral Discs

A comprehensive understanding of the cellular heterogeneity and molecular mechanisms underlying the development,homeostasis, and disease of human intervertebral disks（IVDs） remains challenging. Here, the transcriptomic landscape of 108 108 IVD cells was mapped using single-cell RNA sequencing of three main compartments from young and adult healthy IVDs, including the nucleus pulposus（NP）, annulus fibrosus, and cartilage endplate（CEP）. The chondrocyte subclusters were classified based on their potential regulatory, homeostatic, and effector functions in extracellular matrix（ECM） homeostasis. Notably, in the NP, a PROCR+resident progenitor population showed enriched colony-forming unit-fibroblast（CFU-F） activity and trilineage differentiation capacity. Finally, intercellular crosstalk based on signaling network analysis uncovered that the PDGF and TGF-βcascades are important cues in the NP microenvironment. In conclusion, a single-cell transcriptomic atlas that resolves spatially regulated cellular heterogeneity together with the critical signaling that underlies homeostasis will help to establish new therapeutic strategies for IVD degeneration in the clinic.