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Viability, growth kinetics and stem cell markers of single and clustered cells in human intervertebral discs: implications for regenerative therapies

Abstract

Purpose

There is much interest in the development of a cellular therapy for the repair or regeneration of degenerate intervertebral discs (IVDs) utilising autologous cells, with some trials already underway. Clusters of cells are commonly found in degenerate IVDs and are formed via cell proliferation, possibly as a repair response. We investigated whether these clusters may be more suitable as a source of cells for biological repair than the single cells in the IVD.

Methods

Discs were obtained at surgery from 95 patients and used to assess the cell viability, growth kinetics and stem or progenitor cell markers in both the single and clustered cell populations.

Results

Sixty-nine percent (±15) of cells in disc tissue were viable. The clustered cell population consistently proliferated more slowly in monolayer than single cells, although this difference was only significant at P0–1 and P3–4. Both populations exhibited progenitor or notochordal cell markers [chondroitin sulphate epitopes (3B3(−), 7D4, 4C3 and 6C3), Notch-1, cytokeratin 8 and 19] via immunohistochemical examination; stem cell markers assessed with flow cytometry (CD73, 90 and 105 positivity) were similar to those seen on bone marrow-derived mesenchymal stem cells.

Conclusions

These results confirm those of previous studies indicating that progenitor or stem cells reside in adult human intervertebral discs. However, although the cell clusters have arisen via proliferation, there appear to be no greater incidence of these progenitor cells within clusters compared to single cells. Rather, since they proliferate more slowly in vitro than the single cell population, it may be beneficial to avoid the use of clustered cells when sourcing autologous cells for regenerative therapies.

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Acknowledgements

The Notch-1 (bTAN20) antibody developed by S Artavanis-Tsakonas was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by The University of Iowa, Department of the Biology, Iowa City, IA 52242. We are grateful to the Institute of Orthopaedics Ltd, Oswestry and The Henry Smith Charity for funding, all the spinal surgeons at RJAH Orthopaedic Hospital and Mrs. Annie Kerr for assistance with obtaining patient samples.

Conflict of interest

None.

Author information

Correspondence to Sarah Turner.

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Turner, S., Balain, B., Caterson, B. et al. Viability, growth kinetics and stem cell markers of single and clustered cells in human intervertebral discs: implications for regenerative therapies. Eur Spine J 23, 2462–2472 (2014). https://doi.org/10.1007/s00586-014-3500-y

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Keywords

  • Intervertebral disc
  • Cell therapy
  • Clustered cells
  • Progenitor or stem cells
  • MSCs