Insulin-Transferrin-Selenium Prevent Human Chondrocyte Dedifferentiation And Promote The Formation Of High Quality Tissue Engineered Human Hyaline Cartilage

Abstract

This study was to investigate the effects of Insulin-Transferrin-Selenium (ITS) on the proliferation and quantitative gene expression of adult human nasal septum chondrocytes in monolayer culture expansion and the formation of tissue engineered hyaline cartilage. Effects of ITS on human nasal septum chondrocytes monolayer culture expansion and gene expression were evaluated in various culture media either added with 2% fetal bovine serum (FBS) or 1ng/mL basic fibroblast growth factor plus 1ng/mL transforming growth factor or both serum and growth factors supplementation in comparison with medium added with 10%FBS. Chondrocytes cultured in medium added with 2% fetal bovine serum and growth factors either supplemented with or without ITS were then mixed with pluronic F-127 hydrogel for in vivo tissue engineered cartilage formation in nude mice model. Engineered tissues were removed after 8 weeks of implantation and evaluated with histological staining, immunohistochemistry, transmission electron microscopy and quantitative gene expression analysis. ITS promoted human chondrocytes proliferation and reduced chondrocytes dedifferentiation in media supplemented with serum and growth factors. ITS with 2% FBS and growth factors provided 15-fold increased in chondrocytes number by the end of the culture period compared to the standard culture medium used in chondrocytes culture (medium added with 10% FBS). Engineered tissue resulted from ITS supplementation demonstrated higher quality of cartilage formation. In conclusion, our study has demonstrated the benefits of ITS supplementation in human chondrocytes monolayer culture and tissue engineering cartilage formation.

Effects of co‐cultures of meniscus cells and articular chondrocytes on PLLA scaffolds

“…The serum-free formulation obviates concerns of possible immune rejection and batch-to-batch variations in animal serum (Mandl et al, 2002;Yaeger et al, 1997). This cocktail has been used previously for both meniscus and articular cartilage tissue engineering to maintain cell phenotype and increase proteoglycan synthesis (Chua et al, 2005;Giannoni et al, 2005;Hoben and Athanasiou, 2008;Yates et al, 2005). …”

Effect of Serum and Insulin Modulation on the Organization and Morphology of Matrix Synthesized by Bovine Corneal Stromal Cells

“…41 ). Insulin-transferrinselenium (ITS) is a precisely defined medium supplement that enhances the proliferation and differentiation of several cell types, [42][43][44][45][46][47][48] increases the contraction of collagen gels by rat cardiac fibroblasts, 49 and the synthesis of collagen by avian tendon cells. 44 However, elimination of serum by replacement with insulin or ITS is often detrimental to cell survival, differentiation, and=or proliferation.…”

Creating a spectrum of fibrocartilages through different cell sources and biochemical stimuli

“…FBS, although largely undefined, enhances fibrochondrocyte and chondrocyte growth, but in recent years a ''chondrogenic'' chemically defined medium Mackay et al, 1998;Yoo et al, 1998) has been used to increase proteoglycan synthesis (Adkisson et al, 2001;Chua et al, 2005;Fortier et al, 1997;Giannoni et al, 2005;Schwartz et al, 1974;Yates et al, 2005) and may lend itself to producing the more cartilaginous portion of the fibrocartilage spectrum. This serum-free medium uses insulin-transferrin-selenium (ITSþ) and dexamethasone instead of FBS, and the effects of the insulin in ITSþ, as well as the absence of the many undefined growth factors and other proteins normally found in FBS, may result in profoundly different effects on the growth of cell-based constructs.…”