The Journal of Bone and Joint Surgery, Vol 66, Issue 2 253-259, Copyright © 1984 by Journal of Bone and Joint Surgery, Inc
Studies on cryopreservation of articular cartilage chondrocytes
WW Tomford, GR Fredericks and HJ Mankin
We used cartilage cells isolated from bovine articular cartilage in
experiments to: (1) determine the toxicity of cryopreservatives (glycerol
and dimethyl sulphoxide) on chondrocytes, (2) evaluate methods of freezing
chondrocytes to maximize viability after freezing, and (3) examine the
biosynthetic activity of frozen and thawed chondrocytes in culture. Results
showed that the toxicity of cryopreservatives to chondrocytes is dependent
on the time and temperature of exposure as well as on the concentration of
the cryopreservative. Maximum viability was obtained by a two-stage
freezing procedure using a slow cooling period initially, with
equilibration of the cells at -40 degrees Celsius before further rapid
freezing to -80 degrees Celsius. After seventy-two hours in culture,
chondrocytes that had been frozen using this protocol synthesized products
that appeared by column chromatography to form proteoglycan aggregates.
Clinical Relevance: One of the reasons for failure of frozen osteochondral
allografts is the deterioration of joint function after transplantation due
to degeneration of the articular cartilage. An important factor in the
survival of these cartilage grafts may be preservation of the viability of
the chondrocytes during storage and maintenance of the cell's ability to
function following storage. In this study we evaluated the ability to store
chondrocytes in a frozen state with the aid of cryopreservatives. The
results confirmed that chondrocytes will survive freezing and remain
capable of functioning in the same manner as fresh chondrocytes. This
suggests that chondrocytes in articular cartilage should be able to survive
freezing. The pursuit of methods of preserving articular cartilage by
freezing appears to be warranted.
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