In vitro cartilage formation of composites of synovium-derived mesenchymal stem cells with collagen gel
In vitro cartilage formation of composites of synovium-derived mesenchymal stem cells with collagen gel
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DOI：
10.1007/s00441-005-0010-6
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发表时间：
2005-11-01
影响因子：
3.6
通讯作者：
Muneta, T
中科院分区：
文献类型：
--
作者：
Yokoyama, A;Sekiya, I;Muneta, T
移植物植入是修复软骨缺损最流行的手术之一;然而，牺牲供区一直是一个问题。间充质干细胞（MSC）是再生医学的一个迷人的来源，因为它们可以以侵入性较小的方式收获，易于分离和扩增，具有多潜能，包括软骨形成。MSC可以从包括滑膜的各种成体间充质组织中分离。在这里，我们试图在体外从滑膜来源的骨髓间充质干细胞与胶原凝胶的复合物形成软骨。培养21天后，甲苯胺蓝染色和II型胶原免疫组化显示，复合材料的软骨基质增加。凝胶中由5 × 10（7）和10（8）个细胞/ml组成的复合物比由较低细胞密度组成的复合物更富含蛋白聚糖。1天后，MSC/凝胶复合物收缩，直径减小30%;然而，此后它们是稳定的。通过透射电子显微镜观察到复合材料中具有短突起的圆形细胞产生与软骨细胞相似的形态的胶原纤维。在复合培养过程中，硫酸软骨素和软骨相关基因的mRNA表达增加，表明软骨成熟。使用优化的方法，我们获得了直径为7 mm，厚度为500 μ m的软骨盘。因此，我们的程序应该有可能在体外产生一个大的软骨基质。滑膜来源的骨髓间充质干细胞与胶原凝胶体外复合的自体软骨组织工程移植可能是未来软骨缺损患者移植物植入的替代方法。
Graft implantation is one of the more popular procedures for repairing cartilage defects; however, sacrifices of the donor site have been an issue. Mesenchymal stem cells (MSCs) are a fascinating source for regenerative medicine because they can be harvested in a less invasive manner and are easily isolated and expanded, with multipotentiality including chondrogenesis. MSCs can be isolated from various adult mesenchymal tissues including synovium. Here, we attempted to form cartilage from the composites of synovium-derived MSCs with collagen gel in vitro. After 21 days of culture, the composites had increased their cartilage matrix, as demonstrated by toluidine blue staining and immunohistochemistry for type II collagen. The composites consisting of 5x10(7) and 10(8) cells/ml in gel were richer in proteoglycans than those consisting of lower cell densities. After 1 day, MSCs/gel composites contracted and the diameter decreased by 30%; however, they were stable thereafter. Round cells with short processes producing collagen fibrils showing a similar morphology to that of chondrocytes were seen in the composites by transmission electron microscopy. During composite culture, chondroitin sulfate and mRNA expression for cartilage-related genes increased, demonstrating cartilage maturation. Using an optimized method, we obtained cartilage discs with a diameter of 7 mm and a thickness of 500 mu m. Our procedure should thus make it possible to produce a large cartilage matrix in vitro. The tissue engineering of autologous cartilage from the composites of synovium-derived MSCs with collagen gel in vitro for transplantation may be a future alternative to graft implantation for patients with cartilage defects.