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Collagen type I and decorin expression in tenocytes depend on the cell isolation method

机译:肌腱细胞中I型胶原和核心蛋白的表达取决于细胞分离方法

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Backround The treatment of rotator cuff tears is still challenging. Tendon tissue engineering (TTE) might be an alternative in future. Tenocytes seem to be the most suitable cell type as they are easy to obtain and no differentiation in vitro is necessary. The aim of this study was to examine, if the long head of the biceps tendon (LHB) can deliver viable tenocytes for TTE. In this context, different isolation methods, such as enzymatic digestion (ED) and cell migration (CM), are investigated on differences in gene expression and cell morphology. Methods Samples of the LHB were obtained from patients, who underwent surgery for primary shoulder arthroplasty. Using ED as isolation method, 0.2% collagenase I solution was used. Using CM as isolation method, small pieces of minced tendon were put into petri-dishes. After cell cultivation, RT-PCR was performed for collagen type I, collagen type III, decorin, tenascin-C, fibronectin, Scleraxis, tenomodulin, osteopontin and agreccan. Results The total number of isolated cells, in relation to 1?g of native tissue, was 14 times higher using ED. The time interval for cell isolation was about 17 hours using ED and approximately 50?days using CM. Cell morphology in vitro was similar for both isolation techniques. Higher expression of collagen type I could be observed in tenocyte-like cell cultures (TLCC) using ED as isolation method (p? Conclusion In summary tenocyte-like cells can be obtained with both isolation methods (ED and CM) from the LHB. As no obvious disadvantage could be seen using ED, this method is more suitable for clinical use, as time for cell isolation is shorter and a remarkably higher number of cells can be obtained. However, both isolation methods can further be improved.
机译:背景肩袖撕裂的治疗仍然具有挑战性。肌腱组织工程(TTE)可能会在将来成为替代方案。肌腱细胞似乎是最合适的细胞类型,因为它们易于获得并且无需体外分化。这项研究的目的是检查二头肌腱(LHB)的长头是否可以为TTE输送活的肌腱细胞。在这种情况下,对基因表达和细胞形态的差异研究了不同的分离方法,例如酶促消化(ED)和细胞迁移(CM)。方法从接受原发性肩关节置换手术的患者中获取LHB样本。使用ED作为分离方法,使用0.2%胶原酶I溶液。使用CM作为隔离方法,将碎小筋腱放入培养皿中。细胞培养后,对I型胶原蛋白,III型胶原蛋白,decorin,tenascin-C,纤连蛋白,Scleraxis,tenomodulin,骨桥蛋白和agreccan进行RT-PCR。结果使用ED,相对于1微克天然组织,分离的细胞总数高出14倍。使用ED进行细胞分离的时间间隔约为17小时,使用CM约为50天。两种分离技术的体外细胞形态相似。使用ED作为分离方法,可以在肌腱样细胞培养物(TLCC)中观察到I型胶原的更高表达(p?结论)总之,通过LHB的两种分离方法(ED和CM)均可获得肌腱样细胞。使用ED没有发现明显的缺点,该方法更适合临床使用,因为细胞分离的时间更短并且可以得到明显更多的细胞,但是,两种分离方法都可以进一步改进。

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