Medical Biotechnology Department, Semnan University of Medical Sciences, Semnan, Iran.
Department of Stem Cell and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, P.O. Box: 19395-4644, Tehran, Iran.
Stem Cell Rev Rep. 2018 Oct;14(5):755-766. doi: 10.1007/s12015-018-9816-y.
Growth factors have a pivotal role in chondrogenic differentiation of stem cells. The differential effects of known growth factors involved in the maintenance and homeostasis of cartilage tissue have been previously studied in vitro. However, there are few reported researches about the interactional effects of growth factors on chondrogenic differentiation of stem cells. The aim of this study is to examine the combined effects of four key growth factors on chondrogenic differentiation of mesenchymal stem cells (MSCs). Isolated and expanded rabbit bone marrow-derived MSCs underwent chondrogenic differentiation in a micromass cell culture system that used a combination of the following growth factors: transforming growth factor beta 1 (TGF-β1), bone morphogenetic protein 2 (BMP2), parathyroid hormone related protein (PTHrP), and fibroblast growth factor 2 (FGF2) according to a defined program. The chondrogenic differentiation program was analyzed by histochemistry methods, quantitative RT-PCR (qRT-PCR), and measurement of matrix deposition of sulfated glycosaminoglycan (sGAG) and collagen content at days 16, 23, and 30. The results showed that the short-term combination of TGF-β1 and BMP-2 increased sGAG and collagen content, Alkaline phosphates (ALP) activity, and type X collagen (COL X) expression. Application of either PTHrP or FGF2 simultaneously decreased TGF-β1/BMP-2 induced hypertrophy and chondrogenic markers (at least for FGF2). However, successive application of PTHrP and FGF2 dramatically maintained the synergistic effects of TGF-β1/BMP-2 on the chondrogenic differentiation potential of MSCs and decreased unwanted hypertrophic markers. This new method can be used effectively in chondrogenic differentiation programs.
生长因子在干细胞的软骨分化中起着关键作用。已知在维持和软骨组织内稳态中起作用的生长因子的差异效应已在体外进行了研究。然而，关于生长因子对干细胞软骨分化的相互作用效应的报道很少。本研究旨在研究四种关键生长因子对骨髓间充质干细胞（MSCs）软骨分化的协同作用。分离和扩增的兔骨髓源性 MSCs 在微团细胞培养系统中进行软骨分化，该系统使用以下生长因子的组合：转化生长因子β 1（TGF-β1）、骨形态发生蛋白 2（BMP2）、甲状旁腺激素相关蛋白（PTHrP）和纤维母细胞生长因子 2（FGF2），根据定义的方案。通过组织化学方法、定量 RT-PCR（qRT-PCR）和硫酸化糖胺聚糖（sGAG）和胶原蛋白含量的测量分析软骨分化方案，在第 16、23 和 30 天。结果表明，TGF-β1 和 BMP-2 的短期组合增加了 sGAG 和胶原蛋白含量、碱性磷酸酶（ALP）活性和型 X 胶原（COL X）表达。单独应用 PTHrP 或 FGF2 同时降低了 TGF-β1/BMP-2 诱导的肥大和软骨形成标志物（至少对于 FGF2）。然而，PTHrP 和 FGF2 的连续应用可显著维持 TGF-β1/BMP-2 对 MSCs 软骨分化潜能的协同作用，并降低不必要的肥大标志物。这种新方法可有效用于软骨分化方案。
