The effect of TGF-BETA1, TGF-BETA2, and FGF2 growth factors on the chrondrogenesis of mesenchymal stem cells

Objective: TGFβ1, TGFβ2, and FGF2 growth factors are highly effective in the in vitro chondrogenesis of mesenchymal stem cells. The aim of this study is to investigate the effects of certain days and combinations of TGFβ1, TGFβ2, and FGF2 growth factors on the chondrogenesis process of Adipose-Derived Mesenchymal Stem Cells (ADSCs). Method: ADSCs were cultured in monolayer culture system for 3 days as group I-ITS, group II-ITS, group III-FGF2+TGFβ1, and group IV- FGF2+TGFβ1. After 3 days, the medium was changed, and monolayer culture system was continued for 3 days as group I-ITS, group II-TGFβ2, group III-ITS, and group IV- TGFβ2. ADSCs were treated in 3D pellet culture system for 3 days, as group I-ITS, group II-ITS, group III-FGF2+TGFβ1, group IV-FGF2+TGFβ1, group V-Chondrogenic Medium, and group IV-DMEM+%10FBS. After 3 days, the medium was changed and this time groups were formed as group I-ITS, group II-TGFβ2, group III-ITS, group IV- TGFβ2, group V-chondrogenic medium, and group VI-DMEM+FBS. After 6 days, all groups of the 3D pellet culture system were cultured with a chondrogenic medium for 21 days. Alcian blue staining and Dimethylmethylene Blue (DMMB) analysis were established on the 6th day of monolayer culture systems. In 3D pellet culture systems, we performed alcian blue staining and Dimethylmethylene Blue (DMMB) analysis besides were determined COL2A1- ACAN expressions by immunohistochemistry method. Results: In both monolayer culture and 3D pellet culture systems, which are models of ADSCs chondrogenesis, the combined use of TGFβ1, TGFβ2 and FGF2 had a positive effect on the formation of the cartilage matrix. Conclusion: The application of TGFβ1, TGFβ2, and FGF2 of certain durations and combinations to ADSCs culture system positively supports the in vitro chondrogenesis model. Keywords: ADSCs, TGFβ1, TGFβ2, FGF2 in vitro chondrogenesis