Trophic Effects of Mesenchymal Stem Cells Increase Chondrocyte Proliferation and Matrix Formation
Trophic Effects of Mesenchymal Stem Cells Increase Chondrocyte Proliferation and Matrix Formation
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DOI：
10.1089/ten.tea.2010.0517
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发表时间：
2011-05-01
影响因子：
4.1
通讯作者：
Karperien, Marcel
中科院分区：
文献类型：
--
作者：
Wu, Ling;Leijten, Jeroen C. H.;Karperien, Marcel
以往的研究表明，原代软骨细胞（PC）与各种来源的多能细胞的共培养结果在更高的相对量的软骨基质形成比只含有软骨细胞的文化。本研究的目的是探讨这一观察结果的机制。我们使用了人间充质干细胞（hMSCs）和人PC或牛PC（bPC）的共培养颗粒模型，并研究了在共培养过程中单个细胞群的命运和对软骨形成的贡献。增强的软骨基质沉积通过组织学和总糖胺聚糖沉积的定量来证实。种属特异性定量聚合酶链反应表明，软骨基质基因表达主要来自牛源时，bPC的使用。短串联重复序列分析和物种特异性定量聚合酶链反应分析的基因组DNA证明了近完全丢失的MSC在共培养4周后的培养颗粒。在永生化骨髓间充质干细胞和骨髓基质细胞的共培养颗粒，软骨细胞增殖增加，这是部分模仿使用条件培养基，同时诱导永生化骨髓间充质干细胞的优先凋亡。总之，我们的数据清楚地表明，在MSC和PC的颗粒共培养物中，前者的细胞随着时间的推移而消失。在这些共培养物中增加的软骨形成主要是由于MSC在刺激软骨细胞增殖和软骨细胞基质沉积中的营养作用，而不是MSC积极地经历软骨形成分化。
Previous studies showed that coculture of primary chondrocytes (PCs) with various sources of multipotent cells results in a higher relative amount of cartilage matrix formation than cultures containing only chondrocytes. The aim of this study was to investigate the mechanism underlying this observation. We used coculture pellet models of human mesenchymal stem cells (hMSCs) and human PCs or bovine PCs (bPCs) and studied the fate and the contribution to cartilage formation of the individual cell populations during coculture. Enhanced cartilage matrix deposition was confirmed by histology and quantification of total glycosaminoglycan deposition. Species-specific quantitative polymerase chain reaction demonstrated that cartilage matrix gene expression was mainly from bovine origin when bPCs were used. Short tandem repeat analysis and species-specific quantitative polymerase chain reaction analysis of genomic DNA demonstrated the near-complete loss of MSCs in coculture pellets after 4 weeks of culture. In coculture pellets of immortalized MSCs and bPCs, chondrocyte proliferation was increased, which was partly mimicked using conditioned medium, and simultaneously preferential apoptosis of immortalized MSCs was induced. Taken together, our data clearly demonstrate that in pellet cocultures of MSCs and PCs, the former cells disappear over time. Increased cartilage formation in these cocultures is mainly due to a trophic role of the MSCs in stimulating chondrocyte proliferation and matrix deposition by chondrocytes rather than MSCs actively undergoing chondrogenic differentiation.