In vitro chondrogenesis of human synovium-derived mesenchymal stem cells: Optimal condition and comparison with bone marrow-derived cells
In vitro chondrogenesis of human synovium-derived mesenchymal stem cells: Optimal condition and comparison with bone marrow-derived cells
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DOI：
10.1002/jcb.20546
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发表时间：
2006-01-01
影响因子：
4
通讯作者：
Muneta, T
中科院分区：
文献类型：
--
作者：
Shirasawa, S;Sekiya, I;Muneta, T
越来越多的报道表明间充质干细胞（mesenchymal stern cells，MSCs）存在于骨髓以外的多种组织中，包括滑膜。在这里，我们研究了体外软骨形成的人滑膜来源的MSCs的最佳条件，并比较这些细胞与骨髓来源的MSC，特别是在其软骨形成的潜力。在膝关节韧带损伤手术期间，从六名供体中收获滑膜和骨髓。消化的滑膜细胞或来自骨髓的有核细胞进行克隆扩增。颗粒培养系统用于软骨形成，并评估了七种细胞因子中最多三种细胞因子的最佳组合。以较低密度接种的滑膜来源的MSC扩增得更快。与以前的报道相反，TGF β和地塞米松的组合不足以诱导软骨形成。然而，将BMP 2添加到TGF β和地塞米松中显著增加了软骨颗粒尺寸和软骨基质的合成。还通过电子显微镜和免疫组织学分析了软骨颗粒。在软骨形成过程中，每个颗粒的DNA含量降低，表明颗粒通过新合成的细胞外基质的积累而增加了其大小。通过RT-PCR证实软骨形成基因的顺序表达。滑膜来源的MSC在其表面表位和增殖潜力方面看起来与骨髓来源的MSC相似;然而，在患者匹配的比较中，来自滑膜的软骨颗粒显著大于来自骨髓的软骨颗粒。我们证明了TGF β、地塞米松和BMP 2的组合对于滑膜来源的MSC的体外软骨形成是最佳的，并且滑膜来源的MSC具有比骨髓来源的MSC更大的软骨形成潜力。
There are increasing reports that mesenchymal stern cells (MSCs) are present in various tissues other than bone marrow, including synovium. Here we investigated the optimal conditions for in vitro chondrogenesis of human synovium-derived MSCs and compared these cells with bone marrow-derived MSCs, especially in terms of their chondrogenesis potential. Synovium and bone marrow were harvested from six donors during knee operations for ligament injuries. Digested synovium cells or nucleated cells from bone marrow were expanded clonally. A pellet culture system was used for chondrogenesis, and the best combination of up to three cytokines of the seven assessed. synovium derived MSCs plated at a lower density expanded more rapidly. Contrary to previous reports, a combination of TGF beta and dexamethasone was not sufficient to induce chondrogenesis. However, addition of BMP2 to TGF beta and dexamethasone dramatically increased cartilage pellet size and the synthesis of cartilage matrix. The cartilage pellets were also analyzed by electron microscopy and immunohistology. DNA content per pellet decreased during chondrogenesis, indicating the pellet increased its size through the accumulation of newly synthesized extracellular matrix. Sequential chondrogenic gene expression was demonstrated by RT-PCR. Synovium-derived MSCs looked similar to the bone marrow-derived MSCs in their surface epitopes and proliferation potential; however, cartilage pellets from synovium were significantly larger than those from bone marrow in patient-matched comparisons. We demonstrated that the combination of TGF beta, dexamethasone, and BMP2 was optimal for in vitro chondrogenesis of synovium-derived MSCs and that the synovium-derived MSCs have a greater chondrogenesis potential than bone marrow-derived MSCs.