In order to optimize the tenogenic differentiation of mesenchymal stem cells (MSCs), researchers should consider various factors. However, this requires testing numerous experimental settings, which is costly and time-consuming. We aimed to assess the differential effects of transforming growth factor beta-3 (TGF-β3) on the tenogenesis of tonsil-derived MSCs (T-MSCs) and bone marrow-derived MSCs (BM-MSCs) using response surface methodology (RSM). Bone marrow and tonsillar tissue were collected from four patients; mononuclear cells were separated and treated with 5 or 10 ng/mL of TGF-β3. A full factorial experimental design with a categorical factor of 0 was employed to study the effect of tension based on T-MSCs. Eighty-four trials were fitted with RSM and then used to obtain mathematical prediction models. Exposure of T-MSCs and BM-MSCs to TGF-β3 increased the expression of scleraxis (SCX), tenomodulin (TNMD), decorin, collagen I, and tenascin C. Expression of most of these factors reached a maximum after 2–3 days of treatment. The model predicted that the values of the tenocyte lineage-related factors assessed would be significantly increased at 2.5 days of culture with 2.7 ng/mL of TGF-β3 for T-MSCs and at 2.3 days of culture regardless of TGF-β3 concentration for BM-MSCs. This study demonstrated that the RSM prediction of the culture time necessary for the tenogenic differentiation of T-MSCs and BM-MSCs under TGF-β3 stimulation was similar to the experimentally determined time of peak expression of tenocyte-related mRNAs, suggesting the potential of using the RSM approach for optimization of the culture protocol for tenogenesis of MSCs.

中文翻译：

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响应曲面法优化扁桃体源间充质干细胞与肌腱细胞谱系相关因子

为了优化间充质干细胞（MSCs）的肌腱分化，研究人员应考虑各种因素。但是，这需要测试大量的实验设置，这既昂贵又费时。我们旨在使用响应表面方法（RSM）评估转化生长因子β-3（TGF-β3）对扁桃体来源的MSC（T-MSC）和骨髓来源的MSC（BM-MSC）的增生作用的差异性。收集了四名患者的骨髓和扁桃体组织。分离单个核细胞并用5或10 ng / mL的TGF-β3处理。采用分类因子为0的全因子实验设计来研究基于T-MSC的张力效应。装有RSM的八十四项试验，然后用于获得数学预测模型。T-MSC和BM-MSC暴露于TGF-β3会增加硬化（SCX），腱糖调节蛋白（TNMD），核心蛋白聚糖，胶原I和腱生蛋白C的表达。大多数这些因子的表达在2-3天后达到最大值治疗。该模型预测，与TGF-MSC的TGF-β3浓度无关，在2.5天的2.7 ng / mLTGF-β3培养的T-MSC和2.3天的培养物中，评估的肌腱细胞谱系相关因子的值将显着增加-MSC。这项研究表明RSM预测TGF-β3刺激下T-MSC和BM-MSC肌腱分化所需的培养时间与实验确定的肌腱细胞相关mRNA的峰值表达时间相似，这表明有潜力使用用RSM方法优化MSC肌腱形成的培养方案。