Combination of IGF-1 and TGFβ-3 promote tenocytes differentiation in serum free conditions in vitro

Abstract: By utilizing fractional factorial design, we cultured human tenocytes in vitro in different concentrations of Fetal Bovine Serum (FBS) and combinations of growth factors (IGF-1 and TGFβ-3) supplemented in α-MEM medium to select the growth factors that support tenocyts’ differentiation without FBS for future tissue engineering purposes. Tenocytes were cultured with α-MEM supplemented with 0%, 1%, 5% and 10% FBS and combinations of different concentrations of IGF-1 and TGFβ-3 selected from previously published articles which have potential effect in tenocytes proliferation and differentiation. The result showed that the tenocytes cultured for 14 days in culture medium supplemented with 0% FBS, 50ng/ml IGF-1 and 10ng/ml TGFβ-3 promoted tenocytes differentiation as shown by collagen formation and real time RT-PCR of the tenocytes differentiation markers. This work shows the first time that tenocytes maintained cell survival over 14 days period under FBS free conditions and differentiation can be induced by specific combination of growth factors supplemented in culture medium. Our findings showed the practicability of manageable in vitro tenocytes differentiation with defined culture medium for future tendon tissue engineering