Abstract
The complexity of skeletal pathologies makes use of in vivo models essential to elucidate the pathogenesis of the diseases; nevertheless, chondrocyte and osteoblast cell lines provide relevant information on the underlying disease mechanisms. Due to the limitations of primary chondrocytes, immortalized cells represent a unique tool to overcome this problem since they grow very easily for several passages. However, in the immortalization procedure the cells might lose the original phenotype; thus, these cell lines should be deeply characterized before their use. We immortalized primary chondrocytes from a Cant1 knock-out mouse, an animal model of Desbuquois dysplasia type 1, with a plasmid expressing the SV40 large and small T antigen. This cell line, based on morphological and biochemical parameters, showed preservation of the chondrocyte phenotype. In addition reduced proteoglycan synthesis and oversulfation of glycosaminoglycan chains were demonstrated, as already observed in primary chondrocytes from the Cant1 knock-out mouse. In conclusion, immortalized Cant1 knock-out chondrocytes maintained the disease phenotype observed in primary cells validating the in vitro model and providing an additional tool to further study the proteoglycan biosynthesis defect. The same approach might be extended to other cartilage disorders.
Highlights
Published: 27 August 2021Chondrocyte is the unique cell type in cartilage and is essential for cartilage formation and function
The other major function of chondrocytes occurs in endochondral ossification, in which chondrocytes go through a process of maturation characterized by type X collagen expression, loss of aggrecan and type II collagen expression and cellular hypertrophy, leading to the formation of many skeletal elements and to longitudinal growth of long bones [3]
Cartilageof Cant1 knock-out and Primary chondrocytes isolated from femoral head cartilage
Summary
Chondrocyte is the unique cell type in cartilage and is essential for cartilage formation and function. These cells are embedded in an extracellular matrix (ECM) consisting of complexes of aggrecan, hyaluronan, and link protein in a collagen fibril network [1]. The other major function of chondrocytes occurs in endochondral ossification, in which chondrocytes go through a process of maturation characterized by type X collagen expression, loss of aggrecan and type II collagen expression and cellular hypertrophy, leading to the formation of many skeletal elements and to longitudinal growth of long bones [3]. Chondrocytes maintain cartilage homeostasis by balanced anabolic and catabolic activities. This equilibrium is disrupted in osteoarthritis, the major degenerative pathology associated with chondrocytes.
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