The Effects of 3D Culture on the Expansion and Maintenance of Nucleus Pulposus Progenitor Cell Multipotency

Overview

Journal JOR Spine

Specialty Orthopedics

Date 2021 Mar 29

PMID 33778405

Citations 16

Authors

Julien Guerrero

Sonja Hackel

Andreas S Croft

Christoph E Albers

Benjamin Gantenbein

Affiliations

Soon will be listed here.

Abstract

Introduction: Low back pain (LBP) is a global health concern. Increasing evidence implicates intervertebral disk (IVD) degeneration as a major contributor. In this respect, tissue-specific progenitors may play a crucial role in tissue regeneration, as these cells are perfectly adapted to their niche. Recently, a novel progenitor cell population was described in the nucleus pulposus (NP) that is positive for Tie2 marker. These cells have self-renewal capacity and multipotency potential. However, extremely low numbers of the NP progenitors limit the feasibility of cell therapy strategies.

Objective: Here, we studied the influence of the culture method and of the microenvironment on the proliferation rate and the differentiation potential of human NP progenitors .

Method: Cells were obtained from human NP tissue from trauma patients. Briefly, the NP tissue cells were cultured in two-dimensional (2D) (monolayer) or three-dimensional (3D) (alginate beads) conditions. After 1 week, cells from 2D or 3D culture were expanded on fibronectin-coated flasks. Subsequently, expanded NP cells were then characterized by cytometry and tri-lineage differentiation, which was analyzed by qPCR and histology. Moreover, experiments using Tie2 and Tie2 NP cells were also performed.

Results: The present study aims to demonstrate that 3D expansion of NP cells better preserves the Tie2 cell populations and increases the chondrogenic and osteogenic differentiation potential compared to 2D expansion. Moreover, the cell sorting experiments reveal that only Tie2 cells were able to maintain the pluripotent gene expression if cultured in 3D within alginate beads. Therefore, our results highly suggest that the maintenance of the cell's multipotency is mainly, but not exclusively, due to the higher presence of Tie2 cells due to 3D culture.

Conclusion: This project not only might have a scientific impact by evaluating the influence of a two-step expansion protocol on the functionality of NP progenitors, but it could also lead to an innovative clinical approach.

Citing Articles

Alginate vs. Hyaluronic Acid as Carriers for Nucleus Pulposus Cells: A Study on Regenerative Outcomes in Disc Degeneration.

Ogasawara S, Schol J, Sakai D, Warita T, Susumu T, Nakamura Y Cells. 2024; 13(23).

PMID: 39682732 PMC: 11639827. DOI: 10.3390/cells13231984.

The proteomic landscape of extracellular vesicles derived from human intervertebral disc cells.

Li L, Al-Jallad H, Sun A, Georgiopoulos M, Bokhari R, Ouellet J JOR Spine. 2024; 7(4):e70007.

PMID: 39507593 PMC: 11538033. DOI: 10.1002/jsp2.70007.

Assessment of Tie2-Rejuvenated Nucleus Pulposus Cell Transplants from Young and Old Patient Sources Demonstrates That Age Still Matters.

Otani Y, Schol J, Sakai D, Nakamura Y, Sako K, Warita T Int J Mol Sci. 2024; 25(15).

PMID: 39125917 PMC: 11312270. DOI: 10.3390/ijms25158335.

Getting to the Core: Exploring the Embryonic Development from Notochord to Nucleus Pulposus.

Ambrosio L, Schol J, Ruiz-Fernandez C, Tamagawa S, Joyce K, Nomura A J Dev Biol. 2024; 12(3).

PMID: 39051200 PMC: 11270426. DOI: 10.3390/jdb12030018.

Functionalization of Ceramic Scaffolds with Exosomes from Bone Marrow Mesenchymal Stromal Cells for Bone Tissue Engineering.

Maevskaia E, Guerrero J, Ghayor C, Bhattacharya I, Weber F Int J Mol Sci. 2024; 25(7).

PMID: 38612634 PMC: 11011713. DOI: 10.3390/ijms25073826.

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