低氧对人牙囊细胞生物学特性的影响

发布时间：2018-05-15 22:34

本文选题：低氧 + 人牙囊细胞　；参考：《华西口腔医学杂志》2017年03期

【摘要】：目的研究低氧对人牙囊细胞(h DFCs)生物学特性的影响。方法利用组织块酶消化法从年轻恒牙中分离培养h DFCs;采用免疫荧光技术检测细胞表面标志物,多向诱导实验检测细胞多向分化潜能;模拟体外低氧微环境,将细胞分为常氧组(20%O2)和低氧组(2%O2),分别对两组细胞行Transwell小室试验检测低氧对细胞迁移的影响,采用CCK-8法检测低氧对细胞增殖的影响。通过实时定量聚合酶链反应(q RT-PCR)和Western blot分别从基因和蛋白水平检测h DFCs多能性相关标志物于不同氧体积分数下的表达;分别对两组细胞进行成骨诱导,q RT-PCR检测成骨相关基因,茜素红染色评估矿化结节的形成。结果 h DFCs具有较强的干细胞特征,具有成骨、成脂及成神经多向分化能力,符合间充质干细胞基本标准,能够满足牙组织工程构建对种子细胞的需求。低氧有利于h DFCs多能性的保持,同时促进了h DFCs的迁移和增殖。h DFCs于低氧中进行诱导时,其成骨分化能力得到增强。结论低氧微环境对维持h DFCs多能性,促进h DFCs增殖、迁移和分化有重要作用。
[Abstract]:Objective to study the effects of hypoxia on the biological characteristics of human dental follicle cells (H DFCs). Methods h DFCs was isolated and cultured from young permanent teeth by tissue block enzyme digestion. Immunofluorescence was used to detect cell surface markers and multidirectional induction test was used to detect the multidirectional differentiation potential of the cells; the hypoxic microenvironment was modeled in vitro, and the cells were divided into the normal oxygen group (20%O2). The effect of hypoxia on cell migration was detected in two groups of cells in two groups of Transwell cells, and the effect of hypoxia on cell proliferation was detected by CCK-8 method. The different oxygen volumes of H DFCs multipotent markers were detected by real-time quantitative polymerase chain reaction (Q RT-PCR) and Western blot, respectively. Two groups of cells were induced to induce osteogenesis, Q RT-PCR was used to detect bone related genes, and alizarin red staining was used to evaluate the formation of mineralized nodules. Results h DFCs had strong stem cell characteristics, with osteogenesis, lipid and neurogenic multidirectional differentiation, which met the basic standard of mesenchymal stem cells and could meet the construction of dental tissue engineering. The demand for seed cells. Hypoxia is beneficial to the retention of H DFCs pluriability, while promoting the migration and proliferation of H DFCs, the osteogenic differentiation ability of.H DFCs is enhanced when.H DFCs is induced in hypoxia. Conclusion hypoxia microenvironment plays an important role in maintaining h DFCs multienergy, promoting h DFCs proliferation, migration and differentiation.

【作者单位】：口腔疾病研究国家重点实验室国家口腔疾病临床研究中心四川大学华西口腔医院创伤与整形外科;口腔再生医学国家地方联合工程实验室华西口腔医院(四川大学);
【基金】：国家自然科学基金(81271119)~~
【分类号】：R781

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