Alginate biopolymer as a 3-dimentional microenvironment for differentiation of human mesenchymal stem cells to chondrocytes

The aim of this research was to study the potency of Alginate biopolymer as a microenvironment for differentiation of human adipose tissue derived mesenchymal stem cells (hAMSCs) to chondrocytes. For this purpose، human adipose tissue was prepared from health voluntary donors with liposuction and digested by collagenase type ׀. The mesenchymal stem cells cultured in alginate beads in presence of TGF-β1 and ascorbic acid. Alcian blue staining was done to show the differentiation of mesenchymal stem cell to chondrocytes. RT-PCR was done for the evaluation of collagen type II gene expression as a chondrocytic molecular marker. Immuno-cytochemical staining of isolated cells by anti human Vimentin antibody indicated to more than 95% of positive cells as hAMSCs. After induction of chondrogenic differentiation in alginate beads، alcian blue staining showed expression of sulphated proteoglycans in chondrocytes. Positive stained cells in controls and alginate beaded cultured cells indicated to (36. 5% ± 0. 7 and 56. 5 ± 4. 9 at day 3) and (83% ± 2. 8 and 93 ± 2. 8 at day 14) respectively. RT-PCR results indicated to the expression of collagen type II in alginate bead cultured cells and the comparison of results by controls indicated to significant increased expression in alginate bead cultured cells. (43% ± 4. 5 and 70 ± 5 at day 3) and (54% ± 2. 8 and 77 ± 1. 4 at day 14) respectively. In conclusion، our result indicated and supported the using of alginate beads as 3 dimentional culture system in tissue engineering for chondrogenic culture system.