Temporal Dynamic Changes in Synthesis of Chondroitin Sulfate Isomers in Canine Articular Chondrocyte Culture

Sadatoshi MAEDA, Takayoshi MIYABAYASHI, Janet K. YAMAMOTO, Gregory D. ROBERTS, Allan J. LEPINE, Roger M. CLEMMONS
2003 Journal of Veterinary Medical Science  
To investigate temporal dynamic changes in the synthesis of chondroitin 6-sulfate (CS6) and chondroitin 4-sulfate (CS4) in vitro, normal articular cartilage of femoral heads was harvested from three dogs. Chondrocytes were isolated and cultured in alginate microspheres for 21 days. On days 7, 14 and 21, DNA content was quantified by fluorometric assay using Hoechst 33258. On days 14 and 21, proteoglycans were extracted, and the amounts of CS6 and CS4 were quantified after chondroitinase ABC
more » ... ndroitinase ABC digestion using capillary electrophoresis. The DNA content and amounts of CS6 and CS4 increased during the culture period. The amounts of CS6 and CS4 divided by DNA content revealed that the synthesis of CS6 was more up-regulated than CS4. KEY WORDS: capillary electrophoresis, chondrocyte, chondroitin sulfate.
doi:10.1292/jvms.65.1373 pmid:14709831 fatcat:b5nupxwfsvf4vhqval3lbnsh2e