Monitoring collagen transcription by vascular smooth muscle cells in fibrin-based tissue constructs.
Monitoring collagen transcription by vascular smooth muscle cells in fibrin-based tissue constructs.
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监测基于纤维蛋白的组织结构中血管平滑肌细胞的胶原蛋白转录。
DOI：
10.1089/ten.tec.2009.0112
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
Tranquillo,RobertT
中科院分区：
文献类型：
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作者：
Weinbaum,JustinS;Qi,Jie;Tranquillo,RobertT
目前用于测量工程组织中胶原蛋白含量的方法与监测胶原蛋白产生不相容，因为它们需要破坏组织。我们已经实施了一种基于胰蛋白酶的策略来非侵入性地监测胶原蛋白的产生。基于纤维蛋白的组织构建体使用稳定转染有胶原I启动子/荧光素酶转基因的血管平滑肌细胞制成，其胶原含量与对照细胞相当，但可以在体外组织生长期间非侵入性成像以跟踪胶原转录。我们发现，这些细胞在转录水平上报告了I型胶原的产生，以响应生长因子转化生长因子-β1和β-氨基己酸的纤溶抑制作用，这些变化与mRNA和蛋白质水平的变化一致。由于这些细胞立即报告胶原蛋白的变化而不破坏组织，它们将促进使用多种刺激来产生功能性工程组织的构建优化。
Current methods for measuring collagen content in engineered tissues are incompatible with monitoring of collagen production because they require destruction of the tissue. We have implemented a luciferase-based strategy to monitor collagen production noninvasively. Fibrin-based tissue constructs made using vascular smooth muscle cells stably transfected with a collagen I promoter/luciferase transgene developed with collagen content comparable to control cells, but could be imaged noninvasively to follow collagen transcription during tissue growthin vitro. We showed that these cells reported collagen I production at the transcriptional level in response to the growth factor transforming growth factor-β1 and fibrinolytic inhibition by ɛ-aminocaproic acid and that these changes were consistent with changes at the mRNA and protein levels. As these cells report collagen changes instantly and without tissue destruction, they will facilitate construct optimization using multiple stimuli to produce functional engineered tissues.