Eslaminejad Mohamadreza Baghaban, Karimi Negar, Shahhoseini Maryam
Department of Stem Cells and Developmental Biology at Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, P.O. Box 16635-148, Tehran, Iran,
Histochem Cell Biol. 2013 Dec;140(6):623-33. doi: 10.1007/s00418-013-1121-x. Epub 2013 Jul 10.
A study of the cartilage differentiation of mesenchymal stem cells (MSCs) would be of particular interest since one strategy for cell-based treatment of cartilage defects emphasizes the use of cells that are in a differentiated state. The present study has attempted to evaluate the effects of two well-known glycogen synthase kinase-3 inhibitors, including lithium chloride (LiCl) and SB216763 on a human marrow-derived MSC (hMSC) chondrogenic culture. Passaged-3 MSCs were condensed into small pellets and cultivated in the following groups based on the supplementation of chondrogenic medium: transforming growth factor (TGF)-β1, TGF-β1 + LiCl, TGF-β1 + SB216763, TGF-β3, TGF-β3 + LiCl, and TGF-β3 + SB216763. The cultures were maintained for 21 days and then analyzed for expression of Sox9, aggrecan, collagen II, β-catenin, and axin genes. Deposition of glycosaminoglycan (GAG) in the cartilage matrix was also measured for certain cultures. The presence of both LiCl and SB216763 along with TGF-β in the MSC chondrogenic culture led to the up-regulation of cartilage-specific genes. TGF-β3 appeared much better than TGF-β1. Based on our findings, SB216763 was more effective in up-regulation of cartilage-specific genes. These chondrogenic effects appeared to be mediated through the Wnt signaling pathway since β-catenin and axin tended to be up-regulated and down-regulated, respectively. In the culture with SB216763 + TGF-β3, significantly more GAG was deposited (P < 0.05). In conclusion, addition of either SB216763 or LiCl to hMSC chondrogenic culture up-regulates cartilage-specific gene expression and enhances GAG deposition in the culture.
间充质干细胞（MSCs）软骨分化的研究可能会特别引人关注，因为基于细胞治疗软骨缺损的一种策略强调使用处于分化状态的细胞。本研究试图评估两种著名的糖原合酶激酶-3抑制剂，即氯化锂（LiCl）和SB216763对人骨髓来源的间充质干细胞（hMSC）软骨形成培养的影响。传代3次的间充质干细胞被浓缩成小颗粒，并根据软骨形成培养基的添加情况分为以下几组进行培养：转化生长因子（TGF）-β1、TGF-β1 + LiCl、TGF-β1 + SB216763、TGF-β3、TGF-β3 + LiCl和TGF-β3 + SB216763。培养21天，然后分析Sox9、聚集蛋白聚糖、胶原蛋白II、β-连环蛋白和轴蛋白基因的表达。还对某些培养物测量了软骨基质中糖胺聚糖（GAG）的沉积。在间充质干细胞软骨形成培养中，LiCl和SB216763与TGF-β共同存在导致软骨特异性基因上调。TGF-β3似乎比TGF-β1效果好得多。根据我们的研究结果，SB216763在上调软骨特异性基因方面更有效。这些软骨形成作用似乎是通过Wnt信号通路介导的，因为β-连环蛋白和轴蛋白分别倾向于上调和下调。在SB216763 + TGF-β3的培养物中，GAG沉积显著更多（P < 0.05）。总之，在hMSC软骨形成培养中添加SB216763或LiCl可上调软骨特异性基因表达并增强培养物中GAG的沉积。
