摘要
目的探讨低氧培养的人脐带间充质干细胞对巨噬细胞极化的影响。方法采用组织块贴壁法获取人脐带来源的间充质干细胞(h UC-MSCs),分别将细胞置入常氧(氧浓度21%)和低氧(氧浓度5%)条件下培养,通过细胞成脂、成骨分化诱导观察其多向分化能力,Live/death染色检测细胞活性,ELISA法检测其细胞因子蛋白含量。利用Transwell将常氧和低氧培养的h UC-MSCs与脂多糖(IPS)刺激后的巨噬细胞(THP-1)共培养,免疫荧光检测THP-1的极化情况,ELISA检测THP-1分泌炎症因子以及抗炎因子的情况。结果低氧条件下培养的h UC-MSCs具有成脂、成骨的多向分化潜能;Live/death染色结果显示常氧和低氧培养下的h UC-MSCs均具有较高的细胞活性;低氧条件下培养的h UCMSCs中前列腺素E2(PGE2)、吲哚胺2,3-双加氧酶(IDO)表达水平明显高于常氧培养的h UC-MSCs;低氧条件下培养的h UC-MSCs可促进THP-1向M2型巨噬细胞转化,同时炎症因子肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)的表达明显降低,而抗炎因子IL-10的表达明显增强。结论低氧培养的h UC-MSCs可促使THP-1向M2巨噬细胞转化,提高其抗炎能力。
Objective To investigate the effect of human umbilical cord mesenchymal stem cells（h UC-MSCs） on macrophage polarization under hypoxia.Methods hUC-MSCs were obtained by explants adherent culture and cultured under normoxia（21% O2） and hypoxia（5% O2）.The multi-directional dif erentiation of h UC-MSCs was observed by osteogenic and adipogenic dif erentiation induction.Live/death staining was performed to detect the cell viability,and ELISA was executed to detect the protein content in supernatant of hUC-MSCs.Transwell chamber was employed to co-culture the hUC-MSCs cultured under normoxia and hypoxia and macrophage（THP-1） stimulated by lipopolysaccharide（IPS）,then the polarization of THP-1 was detected by immunol uorescence,and the secretions of inl ammatory factor and anti-inl ammatory factor of THP-1 were detected by ELISA.Results h UC-MSCs cultured under hypoxia showed the ability of osteogenic and adipogenic multi-directional dif erentiation.Live/death staining showed the high cell viability of h UC-MSCs cultured under normoxia and hypoxia.The expression levels of prostaglandin E2（PGE2） and indoleamine 2,3-dioxygenase（IDO） were signii cantly higher in the hUC-MSCs cultured under hypoxia than in those cultured under normoxia.h UCMSCs cultured under hypoxia promoted the polarization of THP-1 to M2,obviously reduced the expression of TNF-α and IL-1β,and increased the expression of IL-10 signii cantly.Conclusion h UC-MSCs cultured under hypoxia may promote the polarization of THP-1 to M2 and improve the viability of anti-inl ammatory.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2016年第7期523-527,共5页
Medical Journal of Chinese People's Liberation Army
基金
国家高技术研究发展计划(863计划)项目(2013AA020105)
国家自然科学基金青年基金(81501682)
解放军总医院科技创新苗圃基金(14KMM43)~~
关键词
低氧
间质干细胞
巨噬细胞极化
炎症
hypoxia
mesenchymal stem cells
macrophage polarization
inflammation
