Abstract
Aim: The aim of this study was to investigate whether viable chondrocytes can be isolated and subsequently expanded in culture, from cryopreserved intact human articular cartilage.
Method: Human articular cartilage samples, retrieved from patient undergoing total knee replacement, were cored as 5 mm diameter discs then minced to approximately 0.1 mm3 size pieces. Samples were cryopreserved at the following stages; intact cartilage discs, minced cartilage and chondrocytes immediately after enzymatic isolation. After completing of isolation, cell viability was examined using LIVE/DEAD fluorescent staining. Isolated chondrocytes were then cultured and a cell proliferation assay was performed at day 4, 7, 14, 21 and 28 days.
Results: The results showed that the viability of isolated chondrocytes from control, cryopreserved intact AC discs, minced AC and isolated then frozen samples were 71.84 ± 2.63%, 25.61 ± 2.41%, 31.32 ± 2.47 % and 42.53 ± 4.66% respectively. Isolated chondrocytes from all groups were expanded by following degrees after 28 days of culture; Group A: 10 times, Group B: 18 times, Group C: 106 times, and Group D: 154 times.
Conclusion: We conclude that viable chondrocytes can be isolated from cryopreserved intact human AC and then cultured to expand their number. This method could be employed to patients benefit undergoing autologous chondrocyte implantation.
Correspondence should be addressed to: Tim Wilton, BASK, c/o BOA, The Royal College of Surgeons, 35–43 Lincoln’s Inn Fields, London WC2A 3PE.
