Tanaka Yuya, Hosoyama Tohru, Mikamo Akihito, Kurazumi Hiroshi, Nishimoto Arata, Ueno Koji, Shirasawa Bungo, Hamano Kimikazu
Department of Surgery and Clinical Science, Graduate School of Medicine, Yamaguchi University 1-1-1 Minami-kogushi, Ube, Yamaguchi 755-8505, Japan.
Department of Surgery and Clinical Science, Graduate School of Medicine, Yamaguchi University1-1-1 Minami-kogushi, Ube, Yamaguchi 755-8505, Japan; Center for Regenerative Medicine, Graduate School of Medicine, Yamaguchi University1-1-1 Minami-kogushi, Ube, Yamaguchi 755-8505, Japan.
Am J Transl Res. 2017 Feb 15;9(2):664-673. eCollection 2017.
Cell sheet technology is a promising therapeutic strategy for the treatment of ischemic diseases such as myocardial infarction. We recently developed a novel protocol, termed "hypoxic preconditioning," capable of augmenting the therapeutic efficacy of cell sheets. Following this protocol, the pro-angiogenic and anti-fibrotic activity of cell sheets were enhanced by brief incubation of cell sheets under hypoxic culture conditions. However, the precise molecular mechanism underlying the hypoxic preconditioning of cell sheets is unclear. In the present study, we examined signal transducers in cell sheets to identify those responsive to hypoxic preconditioning, using cardiosphere-derived cell (CDC) sheets. We initially tested whether sheet-like structures were suitable for hypoxic preconditioning by comparing them with individual cells. Hypoxic preconditioning was more effective in sheeted cells than in individual cells. Expression of hypoxia inducible factor-1α (HIF-1α) and mammalian target of rapamycin (mTOR) were induced upon hypoxic preconditioning of cell sheets, as was the phosphoinositide 3-kinase (PI3K)/Akt pathway. In addition, hypoxic preconditioning increased phosphorylation of epidermal growth factor receptor (EGFR) and heat shock protein 60 (HSP60) in CDC sheets. Our findings provide novel insights into the utility of hypoxic preconditioning in cell sheet-based technologies for the treatment of ischemic diseases.
细胞片技术是一种用于治疗诸如心肌梗死等缺血性疾病的有前景的治疗策略。我们最近开发了一种名为“缺氧预处理”的新方案，能够增强细胞片的治疗效果。按照该方案，通过在缺氧培养条件下对细胞片进行短暂孵育，可增强细胞片的促血管生成和抗纤维化活性。然而，细胞片缺氧预处理背后的确切分子机制尚不清楚。在本研究中，我们使用源自心球的细胞（CDC）片，检测了细胞片中的信号转导分子，以确定那些对缺氧预处理有反应的分子。我们首先通过将片状结构与单个细胞进行比较，测试了片状结构是否适合进行缺氧预处理。缺氧预处理对片状细胞比对单个细胞更有效。细胞片进行缺氧预处理后，缺氧诱导因子-1α（HIF-1α）和雷帕霉素靶蛋白（mTOR）的表达以及磷酸肌醇3激酶（PI3K）/Akt信号通路均被诱导。此外，缺氧预处理增加了CDC片中表皮生长因子受体（EGFR）和热休克蛋白60（HSP60）的磷酸化。我们的研究结果为缺氧预处理在基于细胞片的缺血性疾病治疗技术中的应用提供了新的见解。
