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Preparation of Intact Bovine Tail Intervertebral Discs for Organ Culture
Published on: February 2, 2012
Cryopreserved intervertebral disc with injected bone marrow-derived stromal cells: a feasibility study using organ
Samantha C W Chan1, Benjamin Gantenbein-Ritter, Victor Y L Leung
1AO Research Institute, CH-7270 Davos Platz, Switzerland.
The Spine Journal : Official Journal of the North American Spine Society
|February 23, 2010
Summary
Cryopreserved disc cells show 40% viability and survive for 7 days under simulated physiological loading. Injected bone marrow-derived stromal cells (BMSCs) show limited survival but influence gene expression in degenerative disc disease treatment.
Area of Science:
- Regenerative Medicine
- Biomaterials Science
- Orthopedics
Background:
- Cryopreserved allogeneic intervertebral disc transplantation shows promise for degenerative disc disease (DDD) but suffers from continued degeneration due to cell loss.
- Bone marrow-derived stromal cells (BMSCs) show potential in delaying disc degeneration.
Purpose of the Study:
- To assess the in vitro viability of endogenous and injected BMSCs in cryopreserved intervertebral discs under simulated physiological loading.
- To evaluate the impact of BMSC injection on cell viability and gene expression in cryopreserved discs.
Main Methods:
- Cryopreservation of bovine caudal discs at -196°C.
- Injection of PKH-26-labeled BMSCs (1x10^5 or 2.5x10^5) in hydrogel carriers into the nucleus pulposus.
- 7-day culture in a bioreactor under simulated physiological loading.
- Assessment of cell viability, gene expression (RT-PCR), and proteoglycan content.
Main Results:
- 40% of endogenous disc cells survived cryopreservation and maintained viability over 7 days.
- Injected BMSCs showed 60% initial survival, decreasing to 20% after 7 days.
- BMSC implantation did not affect endogenous cell viability but altered gene expression (Col1a2, Mmp-13 upregulation; Col2a1 downregulation) and increased ACAN expression at lower doses.
Conclusions:
- Endogenous cells in cryopreserved discs can survive for 7 days under simulated physiological loading.
- While injected BMSCs have limited survival, they can influence matrix gene expression in cryopreserved discs.
- Further long-term studies are needed to evaluate the influence of injected BMSCs on native disc cells.
