High-throughput, temporal and dose dependent, effect of vitamins and minerals on chondrogenesis

Tissue engineered hyaline cartilage is plagued by poor mechanical properties largely due to inadequate type II collagen expression. Of note, commonly used defined chondrogenic media lack 14 vitamins and minerals, some of which are implicated in chondrogenesis. Type II collagen promoter-driven Gaussia luciferase was transfected into ATDC5 cells to create a chondrogenic cell with a secreted-reporter. The reporter cells were used in an aggregate-based chondrogenic culture model to develop a high-throughput analytic platform. This high-throughput platform was used to assess the effect of vitamins and minerals, alone and in combination with TGF{beta}1, on type II collagen expression. Significant combinatorial effects between vitamins, minerals and TGF{beta}1 in terms of type II collagen expression and metabolism were discovered. An optimal continual supplement of copper and vitamin K in the presence of TGF{beta}1 gave a 2.5-fold increase in collagen expression over TGF{beta}1 supplemented media alone.\n\nSummaryCurrent defined chondrogenic culture media lack several vitamins and minerals. Type II collagen is the quintessential marker of articular hyaline cartilage, and is commonly deficient in engineered tissue. A type II collagen promoter driven secreted luciferase construct has been transduced into ATDC5 cells and used to assess vitamin and mineral effects on chondrogenesis in a high-throughput format.