Gozel Merve, Senkoylu Karya, Kesim Cem, Hasanreisoglu Murat
Koç University Research Center for Translational Medicine, Koç University, Istanbul, Türkiye.
Koç University School of Medicine, Istanbul, Türkiye.
PLoS One. 2025 Aug 4;20(8):e0329672. doi: 10.1371/journal.pone.0329672. eCollection 2025.
This study aims to demonstrate the formation of tunneling tubes (TNTs) between adipose-derived mesenchymal stem cells (AdMSCs) and retinal pigment epithelial cells (RPE-1) and their alterations in response to experimental stress conditions. Serum starvation was employed as a stress condition to induce TNTs between the AdMSC and RPE-1 cells. The presence of TNTs was demonstrated through immunofluorescence microscopy, while scanning electron microscopy was utilized to determine the average thickness. Cell viability was assessed after stress by CellTiter-Glo, and H2DCFH-DA probes evaluated the cells' reactive oxygen species (ROS) levels. Further, JC-1 labelled mitochondrial exchange between cells via TNTs was confirmed by time-lapse imaging. A transmembrane culture system was employed to inhibit TNTs. In this study, we investigated the role of TNTs in facilitating intercellular communication and mitochondrial transfer between AdMSCs and RPE-1 cells under stress. We found that TNT-mediated mitochondrial transfer from AdMSCs to RPE-1 helps to reduce ROS levels and improve cell viability. We demonstrated that direct interaction between AdMSCs and RPE-1 cells was crucial for stress recovery. Co-culture enhanced the viability and sustained the RPE-1 cells' function after stress-induced damage. Mechanical inhibition of TNT formation decreased cell viability and elevated ROS levels, indicating the importance of TNTs in cellular protection. The findings can provide a new perspective on the therapeutic potential of stem cell-based therapy in protecting retinal pigment epithelium cells against stress-induced damage and promoting tissue regeneration.
本研究旨在证明脂肪来源的间充质干细胞（AdMSCs）与视网膜色素上皮细胞（RPE-1）之间隧道管（TNTs）的形成及其在实验应激条件下的变化。采用血清饥饿作为应激条件来诱导AdMSC和RPE-1细胞之间形成TNTs。通过免疫荧光显微镜证实TNTs的存在，同时利用扫描电子显微镜测定其平均厚度。应激后通过CellTiter-Glo评估细胞活力，H2DCFH-DA探针评估细胞的活性氧（ROS）水平。此外，通过延时成像证实了JC-1标记的细胞间通过TNTs进行的线粒体交换。采用跨膜培养系统抑制TNTs。在本研究中，我们研究了TNTs在应激条件下促进AdMSCs和RPE-1细胞间通讯和线粒体转移中的作用。我们发现TNT介导的从AdMSCs到RPE-1的线粒体转移有助于降低ROS水平并提高细胞活力。我们证明了AdMSCs和RPE-1细胞之间的直接相互作用对于应激恢复至关重要。共培养增强了应激诱导损伤后RPE-1细胞的活力并维持其功能。对TNT形成的机械抑制降低了细胞活力并提高了ROS水平，表明TNTs在细胞保护中的重要性。这些发现可为基于干细胞的疗法在保护视网膜色素上皮细胞免受应激诱导损伤和促进组织再生方面的治疗潜力提供新的视角。
