Isolation of viable chondrocytes with proliferative capacity from cryopreserved human articular cartilage

Abstract

Chondrocytes were collected from the stifle joints of four pigs to study the effect of cryopreservation on the chondrogenic potential of chondrocytes. Half of the cells were cryopreserved for 3 months. Polyglycolic acid scaffolds were cultured with fresh or cryopreserved chondrocytes for 4 weeks. Cell morphology and the quality of engineered tissue were evaluated by scanning electron microscopy, histopathology and biochemical methods. More cells attached to scaffolds at 48 h when fresh chondrocytes were seeded. At 4 weeks, the numbers of cells, DNA and collagen II were greater in constructs engineered by fresh cells. However, the collagen II/DNA ratio did not differ between the two groups. More matrix was identified on a scanning electron microscope and by histopathology in the fresh group. Cartilage engineered with cryopreserved chondrocytes may contain less matrix and fewer cells. These findings most likely resulted from a lack of cell attachment on the matrix secondary to cryopreservation. Future studies are needed to further evaluate the mechanism by which cryopreservation may affect chondrocyte attachment.