OBJECTIVETo optimize the culture media by adding the growth factors required to maintain tenocytes survival and promote their differentiation without fetal bovine serum (FBS) supplementation, in order for the approach to be used for any future tendon tissue engineering.

METHODSThe human tenocytes were cultured in α-MEM media by adding FBS at various concentrations and supplementing both insulin-like growth factor 1 (IGF-1) and transforming growth factor β-3 (TGFβ-3). A number of growth factors were selected that could support tenocytes expansion at reduced differentiated state with the minimum FBS. By employing fractional factorial design, different treatment groups went through AlamarBlue(TM) tests to evaluate the cell number growth whilst collagen quantification by real time RT-PCR technique and tenocyte differentiation were also studied.

RESULTSThe tenocytes cultured for 14 days with 0% FBS, 50 ng/ml IGF-1 and 10 ng/ml TGFβ-3 maintained survival over 14 days, the Cell count were 6228.68 ± 43.87. They were higher than the other experimental groups, but less than 10% FBS control group (13 576.74 ± 286.75, t = 41.29, P < 0.05). The tenocytes cultured in the treated group also showed enhanced collagen synthesis ((0.322 ± 0.003) ng, t = 4.13 - 5.93, P < 0.05).

CONCLUSIONThese findings have shown for the first time that human tenocytes could be maintained survival for a long period of time in the culture media without FBS, having this approach a suitable one for tendon tissue engineering.