@article{68870032b06b43278cc83edc83a94e74,
title = "Characterisation of ovine bone marrow-derived stromal cells (oBMSC) and evaluation of chondrogenically induced micro-pellets for cartilage tissue repair in vivo",
abstract = "Abstract: Bone marrow stromal cells (BMSC) show promise in cartilage repair, and sheep are the most common large animal pre-clinical model. Objective: The objective of this study was to characterise ovine BMSC (oBMSC) in vitro, and to evaluate the capacity of chondrogenic micro-pellets manufactured from oBMSC or ovine articular chondrocytes (oACh) to repair osteochondral defects in sheep. Design: oBMSC were characterised for surface marker expression using flow cytometry and evaluated for tri-lineage differentiation capacity. oBMSC micro-pellets were manufactured in a microwell platform, and chondrogenesis was compared at 2%, 5%, and 20% O2. The capacity of cartilage micro-pellets manufactured from oBMSC or oACh to repair osteochondral defects in adult sheep was evaluated in an 8-week pilot study. Results: Expanded oBMSC were positive for CD44 and CD146 and negative for CD45. The common adipogenic induction ingredient, 3-Isobutyl-1-methylxanthine (IBMX), was toxic to oBMSC, but adipogenesis could be restored by excluding IBMX from the medium. BMSC chondrogenesis was optimal in a 2% O2 atmosphere. Micro-pellets formed from oBMSC or oACh appeared morphologically similar, but hypertrophic genes were elevated in oBMSC micro-pellets. While oACh micro-pellets formed cartilage-like repair tissue in sheep, oBMSC micro-pellets did not. Conclusion: The sensitivity of oBMSC, compared to human BMSC, to IBMX in standard adipogenic assays highlights species-associated differences. Micro-pellets manufactured from oACh were more effective than micro-pellets manufactured from oBMSC in the repair of osteochondral defects in sheep. While oBMSC can be driven to form cartilage-like tissue in vitro, the effective use of these cells in cartilage repair will depend on the successful mitigation of hypertrophy and tissue integration.",
keywords = "Bone marrow stromal cells, Cartilage, Chondrogenesis, Differentiation, Hypertrophy, Mesenchymal stem cells, Micro-pellet, Osteoarthritis, Oxygen",
author = "K. Futrega and E. Music and Robey, {P. G.} and S. Gronthos and R. Crawford and S. Saifzadeh and Klein, {T. J.} and Doran, {M. R.}",
note = "Funding Information: The Translational Research Institute (TRI) is supported by Therapeutic Innovation Australia (TIA). TIA is supported by the Australian Government through the National Collaborative Research Infrastructure Strategy (NCRIS) programme. The authors thank the core facilities at the TRI, including the Flow Cytometry Core and the Microscopy Core. The authors thank Dr. Siamak Saifzadeh{\textquoteright}s team at the Medical Engineering Research Facility (MERF, QUT) for assistance in organising and executing this study. The authors would like to thank Professor MingHao Zheng for technical advice, and Orthocell Ltd. (Perth Australia) for donating the Celgro scaffolds used in these studies. MRD, RWC, and TJK gratefully acknowledge project support from the National Health and Medicine Research Council (NHMRC) of Australia (Project Grant APP1083857) and NHMRC Fellowship support of MRD (APP1130013). KF and PGR are supported by the Intramural Research Program of the NIH, NIDCR. Funding Information: The Translational Research Institute (TRI) is supported by Therapeutic Innovation Australia (TIA). TIA is supported by the Australian Government through the National Collaborative Research Infrastructure Strategy (NCRIS) programme. The authors thank the core facilities at the TRI, including the Flow Cytometry Core and the Microscopy Core. The authors thank Dr. Siamak Saifzadeh?s team at the Medical Engineering Research Facility (MERF, QUT) for assistance in organising and executing this study. The authors would like to thank Professor MingHao Zheng for technical advice, and Orthocell Ltd. (Perth Australia) for donating the Celgro scaffolds used in these studies. MRD, RWC, and TJK gratefully acknowledge project support from the National Health and Medicine Research Council (NHMRC) of Australia (Project Grant APP1083857) and NHMRC Fellowship support of MRD (APP1130013). KF and PGR are supported by the Intramural Research Program of the NIH, NIDCR. Funding Information: MRD, RWC, and TJK gratefully acknowledge project support from the National Health and Medicine Research Council (NHMRC) of Australia (Project Grant APP1083857) and NHMRC Fellowship support of MRD (APP1130013). KF and PGR are supported by the Intramural Research Program of the NIH, NIDCR. The funding bodies played no role in the design of the study and collection, analysis, and interpretation of data or writing of the manuscript. ",
year = "2021",
month = dec,
doi = "10.1186/s13287-020-02045-3",
language = "English",
volume = "12",
journal = "Stem Cell Research and Therapy",
issn = "1757-6512",
publisher = "BioMed Central",
number = "1",
}