Related Experiment Video
Updated: Jun 25, 2026
13:37
Preparation of Intact Bovine Tail Intervertebral Discs for Organ Culture
Published on: February 2, 2012
14.5K
Cryopreserving the intact intervertebral disc without compromising viability.
Ward Shalash1, Ryan Forcier1, Adam Z Higgins1
1School of Chemical, Biological and Environmental Engineering Oregon State University Corvallis Oregon USA.
JOR Spine
|August 6, 2024
Summary
Cryopreservation of intervertebral discs (IVDs) is now possible with a novel method accelerating cryoprotective agent (CPA) delivery. This technique ensures high cell viability after frozen storage, overcoming previous barriers in tissue preservation for transplantation and research.
Area of Science:
- Biomedical Engineering
- Regenerative Medicine
- Tissue Engineering
Background:
- Cryopreservation of tissues like intervertebral discs (IVDs) is hindered by the toxicity of cryoprotective agents (CPAs) and the slow rate of their delivery.
- Current limitations in organ transplantation and regenerative therapy research are partly due to the challenges in preserving tissues and organs for extended periods.
- Human IVDs present a significant opportunity for treating back pain and advancing regenerative therapies, but logistical issues in organ acquisition and transport impede their use.
Purpose of the Study:
- To develop and validate a novel cryopreservation protocol for intact intervertebral discs (IVDs).
- To significantly accelerate the delivery of cryoprotective agents (CPAs) into IVDs to mitigate toxicity and cell death.
- To enable long-term storage of IVDs with preserved tissue viability for potential transplantation and research applications.
Main Methods:
- Tested various cryoprotective agent (CPA) solutions on bovine nucleus pulposus cells to identify the least cytotoxic option.
- Utilized Computed Tomography (CT) imaging with CPA contrast enhancement to quantify saturation times in intact bovine IVDs under dynamic loading and swelling conditions.
- Validated the cryopreservation protocol, including one week of frozen storage, by assessing cell viability in multiple IVD regions post-thaw.
Main Results:
- A cryopreservation medium using dimethyl sulfoxide and ethylene glycol demonstrated over 7.5 hours of safety before cytotoxicity.
- A dynamic loading and swelling protocol reduced IVD CPA saturation time from over 3 days to under 5 hours, a more than 20x improvement.
- After one week of cryopreservation, all tested IVD regions exhibited 85% cell viability, comparable to fresh, unfrozen controls.
Conclusions:
- Developed a novel method for rapid CPA delivery into intact IVDs through post-compression swelling, addressing a key challenge in tissue cryopreservation.
- This accelerated CPA permeation enables effective cryopreservation of IVDs with no significant loss in cell viability.
- The optimized protocol facilitates the potential development of transplantation banks for IVDs and enhances their utility in regenerative medicine research.
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