Department of Biology (Zoology Section), Faculty of Science, Hacettepe University, Beytepe, Ankara 06800, Turkey.
Connect Tissue Res. 2012;53(6):485-91. doi: 10.3109/03008207.2012.694511. Epub 2012 Jul 20.
As a result of repeated movement, tendons are functionally open to traumas. According to this situation, tenocytes have already been used for tissue engineering therapies. It has been reported that long-term monolayer (ML) culture of tenocytes may lead to a phenotypic drift within passages. Depending on our previously published work, it is clearly demonstrated that high-density (HD) culture improves cell growth and differentiation of tenocytes. However, it is not yet established if HD favors the differentiated state during long-term culture. Therefore, we compared the differences in gene expression of tendon collagens and tendon markers of tenocytes from long-term ML and HD culture conditions by quantitative, real-time polymerase chain reaction (QRT-PCR) for over a period of 3 weeks. COLI, COLIII, COLV, Scx, and Tnmd were target genes as the major matrix constituents of tendons as well as being involved in matrix integrity and tenocyte phenotype. According to our results, tenocytes in HD culture synthesized less amounts of COLIII, COLV, and Tnmd, and dependent on the investigation time point, higher amounts of Scx. We consider that tenocytes produced in HD culture system may not provide sufficient efficiency during tissue engineering approaches. By the fact that most molecules showed significantly higher expression profiles in ML culture condition, it is suggested that culture and passage in ML should be taken into consideration for further tissue engineering approaches to maintain a phenotype with less amount of drift.
由于反复运动,肌腱容易受到创伤。根据这种情况,已经使用肌腱细胞用于组织工程治疗。据报道,肌腱细胞的长期单层(ML)培养可能会导致传代过程中的表型漂移。根据我们之前发表的工作,显然表明高密度(HD)培养可以促进肌腱细胞的生长和分化。然而,在长期培养中,HD 是否有利于分化状态尚不确定。因此,我们通过定量实时聚合酶链反应(QRT-PCR)比较了长期 ML 和 HD 培养条件下肌腱细胞的肌腱胶原和肌腱标志物的基因表达差异,培养时间超过 3 周。COL1、COL3、COL5、Scx 和 Tnmd 是主要的基质成分的靶基因,以及参与基质完整性和肌腱细胞表型的靶基因。根据我们的结果,HD 培养的肌腱细胞合成的 COL3、COL5 和 Tnmd 较少,而 Scx 的含量则随研究时间点的不同而不同。我们认为,HD 培养系统中产生的肌腱细胞在组织工程方法中可能效率不高。由于大多数分子在 ML 培养条件下表现出明显更高的表达谱,因此建议在进一步的组织工程方法中考虑 ML 培养和传代,以维持表型漂移较小的状态。