Abstract
Introduction
The use of autologous human primary articular chondrocytes (hPACs) for repairing damaged cartilage is the golden standard; however, their 2-dimensional (2D) expansion induces dedifferentiation, making it challenging to create hyaline cartilage. Spinner flasks are efficient for generating cartilage organoids, allowing hPACs to proliferate without dedifferentiation; however, porcine notochordal cell-derived matrix (NCM) is needed for aggregation, limiting clinical application. Human articular chondroprogenitor cells (hACPCs) can be expanded many fold while maintaining chondrogenic potential. Therefore, the scalable production of hACPC cartilage organoids without NCM in spinner flasks was investigated in this study.
Methods
hPAC organoids with NCM and hACPC organoids using bone morphogenetic protein 9 (BMP-9) were produced in spinner flasks in 14 days. Thereafter, approximately 20 organoids were fused in low adhesive wells for 21 days. Organoids underwent mechanical testing, and both organoids and fused constructs were evaluated using biochemical, histological, and immunohistochemical analysis.
Results
The hACPCs self-assembled and synthesized abundant extracellular matrix once stimulated with BMP-9. The hPAC and hACPC organoids showed similar mechanical properties, but hACPC organoids and fused constructs showed a more uniform matrix and cell distribution.
Conclusion
The hACPC organoids fused into a neo-hyaline cartilage-like tissue, demonstrating their potential for improved, scalable cartilage tissue repair.
The use of autologous human primary articular chondrocytes (hPACs) for repairing damaged cartilage is the golden standard; however, their 2-dimensional (2D) expansion induces dedifferentiation, making it challenging to create hyaline cartilage. Spinner flasks are efficient for generating cartilage organoids, allowing hPACs to proliferate without dedifferentiation; however, porcine notochordal cell-derived matrix (NCM) is needed for aggregation, limiting clinical application. Human articular chondroprogenitor cells (hACPCs) can be expanded many fold while maintaining chondrogenic potential. Therefore, the scalable production of hACPC cartilage organoids without NCM in spinner flasks was investigated in this study.
Methods
hPAC organoids with NCM and hACPC organoids using bone morphogenetic protein 9 (BMP-9) were produced in spinner flasks in 14 days. Thereafter, approximately 20 organoids were fused in low adhesive wells for 21 days. Organoids underwent mechanical testing, and both organoids and fused constructs were evaluated using biochemical, histological, and immunohistochemical analysis.
Results
The hACPCs self-assembled and synthesized abundant extracellular matrix once stimulated with BMP-9. The hPAC and hACPC organoids showed similar mechanical properties, but hACPC organoids and fused constructs showed a more uniform matrix and cell distribution.
Conclusion
The hACPC organoids fused into a neo-hyaline cartilage-like tissue, demonstrating their potential for improved, scalable cartilage tissue repair.
| Original language | English |
|---|---|
| Journal | Cartilage |
| Volume | XX |
| Issue number | X |
| DOIs | |
| Publication status | E-pub ahead of print - 10 Feb 2025 |
Funding
The authors acknowledge Marieke C. van der Steen, PhD, for her contribution in the METC approval to use redundant articular cartilage tissue. This publication is part of the project LS-NeoCarE (with project number NWA.1389.20.192) of the research program NWA-ORC which is (partly) financed by the Dutch Research Council (NWO).
Keywords
- articular chondroprogenitor cells
- cartilage repair
- chondrocytes
- organoids
- tissue engineering