Department of Gastroenterology and Hepatology, Tokyo Medical and Dental University (TMDU), Tokyo, 113-8519, Japan.
Department of Gastroenterology and Hepatology, Tokyo Medical and Dental University (TMDU), Tokyo, 113-8519, Japan; Center for Stem Cell and Regenerative Medicine, Tokyo Medical and Dental University (TMDU), Tokyo, 113-8519, Japan.
Biochem Biophys Res Commun. 2020 Apr 9;524(3):533-541. doi: 10.1016/j.bbrc.2020.01.108. Epub 2020 Feb 1.
The early-phase wound repair response of the intestinal epithelium is characterized by rapid and organized cell migration. This response is regulated by several humoral factors, including TGF-β. However, due to a lack of appropriate models, the precise response of untransformed intestinal epithelial cells (IECs) to those factors is unclear. In this study, we established an in vitro wound repair model of untransformed IECs, based on native type-I collagen. In our system, IECs formed a uniform monolayer in a two-chamber culture insert and displayed a stable wound repair response. Gene expression analysis revealed significant induction of Apoa1, Apoa4, and Wnt4 during the collagen-guided wound repair response. The wound repair response was enhanced significantly by the addition of TGF-β. Surprisingly, addition of TGF-β induced a set of genes, including Slc28a2, Tubb2a, and Cpe, that were expressed preferentially in fetal IECs. Moreover, TGF-β significantly increased the peak velocity of migrating IECs and, conversely, reduced the time required to reach the peak velocity, as confirmed by the motion vector prediction (MVP) method. Our current in vitro system could be employed to assess other humoral factors involved in IEC migration and could contribute to a deeper understanding of the wound repair potentials of untransformed IECs.
肠上皮的早期伤口修复反应的特征是快速且有序的细胞迁移。这种反应受几种体液因子的调节，包括 TGF-β。然而，由于缺乏适当的模型，未转化的肠上皮细胞（IECs）对这些因子的确切反应尚不清楚。在这项研究中，我们基于 I 型天然胶原建立了未转化的 IEC 体外伤口修复模型。在我们的系统中，IEC 在双室培养插入物中形成均匀的单层，并表现出稳定的伤口修复反应。基因表达分析显示，在胶原引导的伤口修复反应过程中，Apoa1、Apoa4 和 Wnt4 的表达显著增加。TGF-β 的添加显著增强了伤口修复反应。令人惊讶的是，TGF-β 诱导了一组基因的表达，包括 Slc28a2、Tubb2a 和 Cpe，这些基因在胎儿 IEC 中优先表达。此外，TGF-β 显著增加了迁移 IEC 的峰值速度，并且通过运动向量预测（MVP）方法证实，相反地，减少了达到峰值速度所需的时间。我们目前的体外系统可用于评估其他参与 IEC 迁移的体液因子，并有助于更深入地了解未转化的 IEC 的伤口修复潜力。
