Trophic Effects of Mesenchymal Stem Cells in Chondrocyte Co-Cultures are Independent of Culture Conditions and Cell Sources
Trophic Effects of Mesenchymal Stem Cells in Chondrocyte Co-Cultures are Independent of Culture Conditions and Cell Sources
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DOI：
10.1089/ten.tea.2011.0715
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发表时间：
2012-08-01
影响因子：
4.1
通讯作者：
Karperien, Marcel
中科院分区：
文献类型：
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作者：
Wu, Ling;Prins, Henk-Jan;Karperien, Marcel
早些时候，我们已经表明，在软骨细胞和骨髓源性间充质干细胞（BM-MSC）的颗粒共培养物中软骨产量的增加是由于MSC在刺激软骨细胞增殖和基质产生中的营养作用，而不是MSC积极进行软骨分化。这些研究在与MSC的软骨形成分化不相容的培养基中进行。在这项研究中，我们测试了MSC的营养作用是否依赖于在与MSC的软骨分化相容的培养基中培养共培养颗粒。此外，我们研究了MSC的营养作用是否依赖于它们的来源，或者是MSC的更普遍的特征。将人BM-MSC和牛原代软骨细胞在与MSC的软骨分化相容的培养基中共培养。通过糖胺聚糖（GAG）定量证实基质产生增强。种特异性定量聚合酶链反应表明，软骨基质主要是牛源性的，表明缺乏骨髓间充质干细胞的软骨分化。此外，随着时间的推移，沉淀共培养物被牛细胞过度生长。为了检测来源对MSC营养作用的影响，将从脂肪组织和滑膜中分离的MSC与人原代软骨细胞共培养，并将其活性与作为对照的BM-MSC进行比较。GAG定量再次证实了软骨基质产生的增加，而与MSC的来源无关。EdU染色结合细胞追踪显示在每种条件下软骨细胞增殖增加。无论MSC来源如何，基因组DNA的短串联重复序列分析显示，随着时间的推移，共培养物中的MSC减少。我们的结果清楚地表明，在共培养颗粒，MSC刺激软骨形成由于软骨细胞的营养作用，而不是分化成软骨细胞，无论培养条件或来源。这意味着MSC在共培养物中的营养作用是一种普遍现象，具有用于软骨修复策略的潜在意义。
Earlier, we have shown that the increased cartilage production in pellet co-cultures of chondrocytes and bone marrow-derived mesenchymal stem cells (BM-MSCs) is due to a trophic role of the MSC in stimulating chondrocyte proliferation and matrix production rather than MSCs actively undergoing chondrogenic differentiation. These studies were performed in a culture medium that was not compatible with the chondrogenic differentiation of MSCs. In this study, we tested whether the trophic role of the MSCs is dependent on culturing co-culture pellets in a medium that is compatible with the chondrogenic differentiation of MSCs. In addition, we investigated whether the trophic role of the MSCs is dependent on their origins or is a more general characteristic of MSCs. Human BM-MSCs and bovine primary chondrocytes were co-cultured in a medium that was compatible with the chondrogenic differentiation of MSCs. Enhanced matrix production was confirmed by glycosaminoglycans (GAG) quantification. A species-specific quantitative polymerase chain reaction demonstrated that the cartilage matrix was mainly of bovine origin, indicative of a lack of the chondrogenic differentiation of MSCs. In addition, pellet co-cultures were overgrown by bovine cells over time. To test the influence of origin on MSCs' trophic effects, the MSCs isolated from adipose tissue and the synovial membrane were co-cultured with human primary chondrocytes, and their activity was compared with BM-MSCs, which served as control. GAG quantification again confirmed increased cartilage matrix production, irrespective of the source of the MSCs. EdU staining combined with cell tracking revealed an increased proliferation of chondrocytes in each condition. Irrespective of the MSC source, a short tandem repeat analysis of genomic DNA showed a decrease in MSCs in the co-culture over time. Our results clearly demonstrate that in co-culture pellets, the MSCs stimulate cartilage formation due to a trophic effect on the chondrocytes rather than differentiating into chondrocytes, irrespective of culture condition or origin. This implies that the trophic effect of MSCs in co-cultures is a general phenomenon with potential implications for use in cartilage repair strategies.