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Updated: Jul 4, 2025
Isolation of Adult Spinal Cord Nuclei for Massively Parallel Single-nucleus RNA Sequencing
Published on: October 12, 2018
Characterization of the Nucleus Pulposus Progenitor Cells via Spatial Transcriptomics
Yu Chen1,2, Long Zhang1,2, Xueqing Shi1,2
1The First Affiliated Hospital of Xiamen University-ICMRS Collaborating Center for Skeletal Stem Cells, State Key Laboratory of Cellular Stress Biology, Faculty of Medicine and Life Sciences, School of Medicine, Xiamen University, Xiamen, 361102, China.
This study reveals that Cathepsin K marks nucleus pulposus progenitor cells (NPPCs) in mice, originating from the NP-outer region. Tie2 is confirmed not to be a marker for NPPCs, clarifying cell origins in intervertebral disc degeneration.
Area of Science:
- Bone biology
- Cellular and molecular biology
- Regenerative medicine
Background:
- Intervertebral disc (IVD) degeneration is linked to nucleus pulposus (NP) cellularity loss.
- The differentiation pathway of NP cells is not fully understood, despite identified NP progenitor cell (NPPC) markers.
- Cell fragility limits transcriptomic analysis of NP-derived cells.
Purpose of the Study:
- To create the first spatially resolved transcriptional atlas of the mouse IVD.
- To identify and characterize NP progenitor cells (NPPCs) and their spatial distribution.
- To validate or refute existing NPPC markers like Tie2.
Main Methods:
- 10x Genomics Visium platform for spatial transcriptomics of mouse IVD.
- Cell lineage tracing experiments.
- In situ sequencing (ISS) analysis.
Main Results:
- NP spots were clustered, revealing NPPC markers like Cathepsin K (Ctsk) are rare and located in the NP-outer subset.
- Cell lineage tracing confirmed Ctsk-expressing cells generate adult NP tissue.
- Tie2 expression was absent in NP subsets and did not label NPPCs, as validated by ISS and cre-mediated lineage tracing.
Conclusions:
- Established the first spatial transcriptomic map of the IVD, providing a valuable resource.
- Identified Ctsk as a marker for NP progenitor cells originating from the NP-outer region.
- Disproven Tie2 as an NPPC marker, correcting a long-held assumption in IVD research.
