The Role of proteoglycans in chondrocyte differentiation of mesenchymal stem cells

蛋白多糖在间充质干细胞软骨细胞分化中的作用

基本信息

批准号：
15590355
负责人：
WATANABE Hideto
金额：
$ 2.3万
依托单位：
Aichi Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590355/
关键词：
cartilage chondrocyte differentiation mesenchymal stem cell TGF-beta proteoglycnn versican PG-M ultrasound BMP TGF-β

项目摘要

1. We evaluated the effects of ultrasound (US) on differentiation of mesenchymal stem cells (MSCs) toward chondrocyte snarl cartilage matrix formation. When human MSCs cultured in the pellets were treated with transforming growth factor-beta (TGF-b), they differentiated into chondrocytes as assessed by alcian blue staining and immunostaining for aggrecan, but non-treated cell pellets and cell pellets treated with BMP-6 alone did not. Furthermore, when low-intensity US was applied for 20 min everyday to the TGF-beta-treated cell pellets, chondrocyte differentiation was enhanced. These results suggest that TGF-b is essential for chondrocyte differentiation of MSCs in pellets and US enhances the differentiation.2. Using chondrogenic/chondrocytic N1511 cells, we evaluated the role of versican/PG-M in chondrocyet differentiation. N1511 cells exhibited transient high expression of versican at an early stage and that of aggrecan at a late stage, which reflects their in vivo expression patterns during chondrocyte differentiation. When versican expression was inhibited using an antisense technology, the following aggrecan expression was decreased and delayed, as assessed by realtime RT-PCR. These results suggest that transient versican expression at the early stage positively regulate chondrocyte differentiaion.3. In addition to high transient expression, versican is continuously expressed, especially in the articular cartilage. We found that versican is colocalized with both hyaluronan and link protein there. Biochemical analysis further confirmed the presence of versican aggregate. These results suggest the presence of tow proteoglycan aggregates, which may play distinct roles.

1. 我们评估了超声 (US) 对间充质干细胞 (MSC) 向软骨细胞软骨基质形成分化的影响。当用转化生长因子-β (TGF-b) 处理沉淀中培养的人 MSC 时，通过阿新蓝染色和蛋白聚糖免疫染色评估，它们分化为软骨细胞，但未经处理的细胞沉淀和用 BMP-6 处理的细胞沉淀独自一人没有。此外，当每天对 TGF-β 处理的细胞沉淀应用 20 分钟低强度超声时，软骨细胞分化得到增强。这些结果表明TGF-b对于颗粒中MSCs的软骨细胞分化至关重要，并且US增强了分化。2．使用软骨形成/软骨细胞 N1511 细胞，我们评估了多功能蛋白聚糖/PG-M 在软骨细胞分化中的作用。 N1511细胞在早期表现出多功能蛋白聚糖的短暂高表达，在晚期表现出聚集蛋白聚糖的短暂高表达，这反映了它们在软骨细胞分化过程中的体内表达模式。通过实时 RT-PCR 评估，当使用反义技术抑制多功能蛋白聚糖表达时，随后的聚集蛋白聚糖表达减少并延迟。这些结果表明早期短暂的多功能蛋白聚糖表达对软骨细胞的分化具有正向调节作用。3．除了高瞬时表达外，多功能蛋白聚糖持续表达，尤其是在关节软骨中。我们发现多功能蛋白聚糖与透明质酸和连接蛋白共定位。生化分析进一步证实了多功能蛋白聚糖聚集体的存在。这些结果表明存在两种蛋白聚糖聚集体，它们可能发挥不同的作用。