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Journal of Bone and Joint Surgery - British Volume, Vol 90-B, Issue 9, 1245-1248.
doi: 10.1302/0301-620X.90B9.20652  
Copyright © 2008 by British Editorial Society of Bone and Joint Surgery
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The viability and proliferation of human chondrocytes following cryopreservation

Z. Xia, DPhil, Research Fellow1; D. Murray, MD, FRCS(Orth), Professor of Orthopaedic Surgery1; P. A. Hulley, PhD, ARC Research Fellow1; J. T. Triffitt, PhD, Professor of Bone Metabolism1; and A. J. Price, DPhil, FRCS(Orth), Reader in Musculoskeletal Science and Consultant Orthopaedic Surgeon1

Botnar Research Centre, Nuffield Department of Orthopaedic Surgery Nuffield Orthopaedic Centre, Headington, Oxford OX3 7LD, UK.

Correspondence should be sent to Mr A. J. Price; e-mail: andrew.price{at}ndos.oc.ac.uk

Human articular cartilage samples were retrieved from the resected material of patients undergoing total knee replacement. Samples underwent automated controlled freezing at various stages of preparation: as intact articular cartilage discs, as minced articular cartilage, and as chondrocytes immediately after enzymatic isolation from fresh articular cartilage. Cell viability was examined using a LIVE/DEAD assay which provided fluorescent staining. Isolated chondrocytes were then cultured and Alamar blue assay was used for estimation of cell proliferation at days zero, four, seven, 14, 21 and 28 after seeding. The mean percentage viabilities of chondrocytes isolated from group A (fresh, intact articular cartilage disc samples), group B (following cryopreservation and then thawing, after initial isolation from articular cartilage), group C (from minced cryopreserved articular cartilage samples), and group D (from cryopreserved intact articular cartilage disc samples) were 74.7% (95% confidence interval (CI) 73.1 to 76.3), 47.0% (95% CI 43 to 51), 32.0% (95% CI 30.3 to 33.7) and 23.3% (95% CI 22.1 to 24.5), respectively. Isolated chondrocytes from all groups were expanded by the following mean proportions after 28 days of culturing: group A ten times, group B 18 times, group C 106 times, and group D 154 times.

This experiment demonstrated that it is possible to isolate viable chondrocytes from cryopreserved intact human articular cartilage which can then be successfully cultured.






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Hip, Knee, Trauma, Upper limb, Foot & Ankle, Paediatrics, Oncology, Spine, Arthroplasty, General