人胎盘胎儿侧间充质干细胞无血清培养上清的抗氧化活性分析  

Anti-oxidative activities of supernatants of human fetal placental mesenchymal stem cells cultured in serum-free medium

作　　者：付雪[1,2] 张玉洁[1,2] 严秀蕊[2] 马晓娜[2] 刘晓明[2] 魏军[1,2] Fu Xue Zhang Yu-jie Yan Xiu-rui Ma Xiao-na Liu Xiao-ming Wei Jun(Clinical Medicine College, Ningxia Medical University, Yinchuan 750003, Ningxia Hui Autonomous Region, China Ningxia Human Stem Ceil Institute, Affiliated Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China)

机构地区：[1]宁夏医科大学临床医学院,宁夏回族自治区银川市750003 [2]宁夏医科大学总医院人类干细胞研究所,宁夏回族自治区银川市750004

出　　处：《中国组织工程研究》2017年第5期773-779,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81460247)~~

摘　　要：背景:目前关于间充质干细胞的研究大多集中于其免疫调节功能,而关于细胞和培养上清的其抗氧化能力的研究较为少见。目的:观察人胎盘胎儿侧间充质干细胞在无血清培养条件下,其上清的抗氧化能力。方法:采用无血清培养基培养胎儿侧胎盘间充质干细胞,分别于48 h收集P2-P6代细胞培养上清。在空白培养基中添加维生素C 100μmol/L作为阳性对照。检测P2-P6代人胎盘胎儿侧间充质干细胞培养上清的总抗氧化能力、清除活性氧自由基的能力和抗氧化酶活性。结果与结论:(1)抗氧化能力:各代次人胎盘胎儿侧间充质干细胞培养上清具有一定的抗氧化能力,且不同代次间的上清具有一定的差异,其中总抗氧化能力与40-80μmol/L的维生素C相当,各代次上清均具有一定的清除二苯基苦基苯肼自由基、羟自由基、超氧阴离子自由基等自由基清除能力;(2)抗氧化酶活性分析:各代次间的人胎盘胎儿侧间充质干细胞培养上清中均可以检测到一定水平的超氧化物歧化酶和谷胱甘肽过氧化物酶活性;(3)结果提示,在无血清条件下培养胎儿侧胎盘间充质干细胞的上清具有一定的抗氧化能力和抗氧化酶的活性。人胎盘胎儿侧间充质干细胞的抗氧化能力与其旁分泌机制有关,但其中的抗氧化成分及分子机制有待进一步的研究。BACKGROUND： Current research on mesenchymal stem cells（MSCs） is mostly focused on its immune regulatory function, while little is reported on the antioxidant capacity of the cells and culture supernatant.OBJECTIVE： To investigate the anti-oxidative capacity of the supernatant harvested from human fetal placenta MSCs（f PMSCs） under a condition of serum free culture. METHODS： f PMSCs were cultured with serum free media, and the supernatants of cells at passages 2-6 were collected at 48 hours after culture. Vitamin C was added into the culture medium, as a positive control, and its concentration was 100 μmol/L. The total antioxidant capacity, scavenging capacity of free radicals and antioxidant enzymatic activities of supernatants were measured. RESULTS AND CONCLUSION： By comparing anti-oxidative activities of vitamin C and na？ve culture medium, supernatants collected from f PMSCs cultures exhibited obvious antioxidant capacities at different extents between passages of cell cultures. The total antioxidant capacity of the culture supernatant was comparable to 40-80 μmol/L vitamin C. In addition, all supernatants derived from cells with different passages displayed capacities to scavenge free radicals, including 2,2-diphenyl-1-picrylhydrazyl radical（DPPH·）, hydroxyl radical（·OH）, superoxide anion radical（O2-）. Even more, activities of antioxidant enzymes, including superoxide dismutase and glutathione peroxidase, were also detected in supernatants collected from different passages of f PMSCs. Under the serum-free condition, the culture supernatants of f PMSCs have antioxidant capacities at certain extent. However, the antioxidant components and underlying mechanisms need to be further studied.

关 键 词：干细胞 组织工程 胎盘 分化 胎儿侧 间充质干细胞 无血清培养 培养上清 抗氧化 自由基清除 抗氧化酶 旁分泌 无细胞治疗 国家自然科学基金 

分 类 号：R394.2[医药卫生—医学遗传学]