Single-cell RNA sequencing reveals the CRTAC1+ population actively contributes to the pathogenesis of spinal ligament degeneration by SPP1+ macrophage

Degenerative spinal stenosis is a chronic disease that affects the spinal ligaments and associated bones, resulting in back pain and disorders of the limbs among the elderly population. There are few preventive strategies for such ligament degeneration. We here aimed to establish a comprehensive transcriptomic atlas of ligament tissues to identify high-priority targets for pharmaceutical treatment of ligament degeneration. Here, single-cell RNA sequencing was performed on six degenerative ligaments and three traumatic ligaments to understand tissue heterogeneity. Tissue samples, including bone (spinous process, facet, and lamina) and ligaments (ligamentum flavum, interspinous ligament, and supraspinal ligament) from Homo Sapiens were obtained during spinal canal decompression surgery. They were then sorted by Fluorescence-activated cell sorting using BD FACS Aria III (BD Biosciences) into CD45+ and CD45-, followed by applying CD45+ and CD45- cells at 3:7 ratio to the 10X Genomics system.