Recapitulating the notochord to nucleus pulposus transition from human mesendoderm progenitors in 3D

Abstract

Intervertebral disc (IVD) degeneration, a major cause of low back pain, is driven by the disruption of nucleus pulposus (NP) tissue homeostasis. The NP arises from embryonic notochord cells that differentiate and adapt to the hypoxic, matrix-rich environment of the developing disc. Recapitulating the developmental trajectory from notochordal progenitors to NP-like cells in vitro remains challenging, as most protocols bypass early specification steps and fail to capture this understudied lineage. Here, using a three-dimensional (3D) system, we reconstruct this developmental path from human pluripotent stem cells into mesendoderm progenitor cells (MEPCs), notochord-like cells (NLCs), and NP-like cells, guided by single-cell transcriptomics and spatial profiling. We identify MEPCs as a distinct progenitor population with a unique gene signature and conserved regulatory networks (GSC, KLF8, LHX1, MIXL1) shared with early organizer and mesendodermal cells. Self-organization in MEPCs-derived aggregates induced spontaneous expression of NOTO, a key transcription factor orchestrating notochordal identity, and enhanced commitment to notochordal fate. Long-term culture in a medium containing TGF-{beta}1 and GDF-5 promoted maturation into SOX9+, KRT8+, CD109+ NP-like cells exhibiting robust matrix deposition and prominent LAMP1-positive intracytoplasmic vacuoles. This scalable, physiologically relevant model enables fundamental studies of human disc biology and early degenerative processes, while advancing regenerative strategies.

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BibTeX RIS

BibTeX

@misc{warin2026recapitulating,
  title = {{Recapitulating the notochord to nucleus pulposus transition from human mesendoderm progenitors in 3D}},
  author = {J. Warin and N. Vedrenne and B. Guidoux and C. Chedeville and M. Blanche and C. Maillard and C. Chariau and C. Dube and R. Desprat and B. Gantenbein and S. M. Richardson and J. Guicheux and L. David and M. A. Tryfonidou and J.-M. LEMAITRE and A. CAMUS},
  howpublished = {bioRxiv},
  year = {2026},
  month = oct,
  doi = {10.64898/2026.10.05.756547},
  url = {https://www.biorxiv.org/content/10.64898/2026.10.05.756547v1},
  note = {CC BY-NC},
}

RIS

TY  - UNPB
TI  - Recapitulating the notochord to nucleus pulposus transition from human mesendoderm progenitors in 3D
AU  - J. Warin
AU  - N. Vedrenne
AU  - B. Guidoux
AU  - C. Chedeville
AU  - M. Blanche
AU  - C. Maillard
AU  - C. Chariau
AU  - C. Dube
AU  - R. Desprat
AU  - B. Gantenbein
AU  - S. M. Richardson
AU  - J. Guicheux
AU  - L. David
AU  - M. A. Tryfonidou
AU  - J.-M. LEMAITRE
AU  - A. CAMUS
PY  - 2026
DA  - 2026/10/05
PB  - bioRxiv
DO  - 10.64898/2026.10.05.756547
UR  - https://www.biorxiv.org/content/10.64898/2026.10.05.756547v1
AB  - Intervertebral disc (IVD) degeneration, a major cause of low back pain, is driven by the disruption of nucleus pulposus (NP) tissue homeostasis. The NP arises from embryonic notochord cells that differentiate and adapt to the hypoxic, matrix-rich environment of the developing disc. Recapitulating the developmental trajectory from notochordal progenitors to NP-like cells in vitro remains challenging, as most protocols bypass early specification steps and fail to capture this understudied lineage. Here, using a three-dimensional (3D) system, we reconstruct this developmental path from human pluripotent stem cells into mesendoderm progenitor cells (MEPCs), notochord-like cells (NLCs), and NP-like cells, guided by single-cell transcriptomics and spatial profiling. We identify MEPCs as a distinct progenitor population with a unique gene signature and conserved regulatory networks (GSC, KLF8, LHX1, MIXL1) shared with early organizer and mesendodermal cells. Self-organization in MEPCs-derived aggregates induced spontaneous expression of NOTO, a key transcription factor orchestrating notochordal identity, and enhanced commitment to notochordal fate. Long-term culture in a medium containing TGF-{beta}1 and GDF-5 promoted maturation into SOX9+, KRT8+, CD109+ NP-like cells exhibiting robust matrix deposition and prominent LAMP1-positive intracytoplasmic vacuoles. This scalable, physiologically relevant model enables fundamental studies of human disc biology and early degenerative processes, while advancing regenerative strategies.
ER  -

Abstract

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