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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（MSCs）向软骨细胞和软骨基质分化的影响。当用转化生长因子- β (TGF-b)处理球培养的人间充质干细胞时，通过阿利新蓝染色和聚集蛋白免疫染色评估，它们分化成软骨细胞，但未处理的细胞球和单独用BMP-6处理的细胞球没有分化成软骨细胞。此外，当低强度US每天作用于tgf - β处理的细胞颗粒20分钟时，软骨细胞分化增强。这些结果表明，TGF-b在骨髓间充质干细胞的软骨细胞分化中起重要作用，而US可促进骨髓间充质干细胞的分化。使用成软骨细胞/软骨细胞N1511，我们评估了versican/PG-M在软骨细胞分化中的作用。N1511细胞在早期表现出一过性的高表达，在后期表现出聚集蛋白的高表达，这反映了它们在软骨细胞分化过程中的体内表达模式。当使用反义技术抑制versican表达时，通过实时RT-PCR评估，以下aggrecan表达降低和延迟。这些结果表明，早期短暂的versican表达对软骨细胞分化具有正向调节作用。除了高瞬时表达外，versican是持续表达的，特别是在关节软骨中。我们发现versican与透明质酸和连接蛋白在那里共定位。生化分析进一步证实了多聚体的存在。这些结果表明存在两种蛋白多糖聚集体，它们可能起着不同的作用。