Electric Field Stimulation of Stem Cell Culture Media Increases Oxygen Radical Uptake Capacity

Maki Yoshikawa, Masatoshi Oshita, T. Gotoh, Y. Tawa, K. Dan
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Abstract

BACKGROUND: Stem cell culture medium is garnering attention because it comprises growth factors and exosomes secreted by stem cells. In this study, stem cell culture medium was subjected to electric field stimulation using the cells alive system (CAS) to determine the antiaging effects of this medium on dermal fibroblasts.METHODS: The medium was added to the monolayer cultures of human fibroblasts with or without 100 μg/mL advanced glycation end products (AGE), and the cells were incubated for 4 hours. Total RNA was extracted, and mRNA levels of several AGE receptors and heat shock proteins were measured using quantitative reverse transcriptase-polymerase chain rection (qRT-PCR).RESULTS: Similar amounts of exosomes were extracted from the supernatants of stem cells cultured with (CAS+) and without (CAS−) electric field stimulation. The results demonstrated that CAS+ had a higher oxygen radical absorbance capacity than CAS−, indicating its high antioxidant activity. Furthermore, the expression of domain-containing link scavenger receptor-1 mRNA, which is involved in the degradation of cell surface receptors for AGE under glycation stress, significantly increased. The electric field stimulation also increased the expression of several heat shock proteins, which play important roles in cellular responses generated against glycation stress. The electric field stimulation of the stem cell culture medium effectively promoted the endocytosis and degradation of AGE, thereby exerting antioxidant effects and inducing glycation stress.CONCLUSION: Electric field stimulation of stem cell culture medium enhanced the expression of Hsp and its antioxidant activity in fibroblasts.KEYWORDS: AGE, electric field stimulation, exosome, fibroblast, oxygen radical absorbance capacity, stem cell
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电场刺激干细胞培养基提高氧自由基吸收能力
背景:干细胞培养基因含有干细胞分泌的生长因子和外泌体而备受关注。在本研究中,使用细胞活系统(CAS)对干细胞培养基进行电场刺激,以确定该培养基对真皮成纤维细胞的抗衰老作用。方法:将培养基加入加或不加100 μg/mL晚期糖基化终产物(AGE)的人成纤维细胞单层培养中,培养4小时。提取总RNA,采用定量逆转录-聚合酶链式反应(qRT-PCR)测定几种AGE受体和热休克蛋白的mRNA水平。结果:在(CAS+)和(CAS−)电场刺激下培养的干细胞上清液中提取了相似数量的外泌体。结果表明,CAS+具有比CAS−更高的氧自由基吸收能力,表明其具有较高的抗氧化活性。此外,糖基化胁迫下参与AGE细胞表面受体降解的结构域链接清除受体-1 mRNA的表达显著增加。电场刺激还增加了几种热休克蛋白的表达,这些蛋白在糖基化应激产生的细胞反应中起重要作用。电场刺激干细胞培养基有效促进AGE的内吞降解,从而发挥抗氧化作用,诱导糖基化应激。结论:电场刺激干细胞培养基可增强成纤维细胞热休克蛋白的表达及其抗氧化活性。关键词:AGE,电场刺激,外泌体,成纤维细胞,氧自由基吸收能力,干细胞
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