National Heart and Lung Institute, Imperial College London, London, UK.
Respiratory & Critical Care Medicine, The University of Hong Kong-Shenzhen Hospital, Shenzhen, Guangdong, People's Republic of China.
Methods Mol Biol. 2021;2269:93-105. doi: 10.1007/978-1-0716-1225-5_7.
Mesenchymal stem cells (MSCs) have emerged as an attractive candidate for cell-based therapy. In the past decade, many animal and pilot clinical studies have demonstrated that MSCs are therapeutically beneficial for the treatment of obstructive lung diseases such as asthma and chronic obstructive pulmonary disease (COPD). However, due to the scarcity of adult human MSCs, human-induced pluripotent stem cells mesenchymal stem cells (iPSCs) are now increasingly used as a source of MSCs. iPSCs are derived by reprogramming somatic cells from a wide variety of tissues such as skin biopsies and then differentiating them into iPSC-MSCs. One of the mechanisms through which MSCs exert their protective effects is mitochondrial transfer. Specifically, transfer of mitochondria from iPSC-MSCs to lung cells was shown to protect lung cells against oxidative stress-induced mitochondrial dysfunction and apoptosis and to reduce lung injury and inflammation in in vivo models of lung disease. In this chapter, we detail our methods to visualize and quantify iPSC-MSC-mediated mitochondrial transfer and to study its effects on oxidant-induced airway epithelial and smooth muscle cell models of acute airway cell injury.
间充质干细胞（MSCs）已成为细胞治疗的有吸引力的候选者。在过去的十年中，许多动物和初步临床研究表明，MSCs 对治疗阻塞性肺部疾病如哮喘和慢性阻塞性肺疾病（COPD）具有治疗益处。然而，由于成人人类 MSCs 的稀缺性，人诱导多能干细胞间充质干细胞（iPSCs）现在越来越多地被用作 MSCs 的来源。iPSCs 通过重编程来自各种组织（如皮肤活检）的体细胞核来获得，然后将其分化为 iPSC-MSCs。MSCs 发挥其保护作用的机制之一是线粒体转移。具体而言，从 iPSC-MSCs 转移线粒体被证明可以保护肺细胞免受氧化应激诱导的线粒体功能障碍和细胞凋亡，并减少肺部损伤和炎症在肺部疾病的动物模型中。在本章中，我们详细介绍了可视化和量化 iPSC-MSC 介导的线粒体转移的方法，并研究了其对氧化剂诱导的气道上皮和平滑肌细胞急性气道细胞损伤模型的影响。
