3T3-L1细胞与人骨髓间充质干细胞体外成脂及脂肪细胞功能的比较研究

发布时间：2018-02-16 12:10

本文关键词： T-L细胞人骨髓间充质干细胞成脂分化　出处：《中国实验血液学杂志》2015年06期 　论文类型：期刊论文

【摘要】：目的:探讨3T3-L1细胞与人骨髓间充质干细胞(mesenchymal stem cells,MSCs)体外成脂及脂肪细胞功能的差异。方法:用密度梯度离心和贴壁培养的方法分离纯化人骨髓MSC,在倒置显微镜下观察细胞形态,体外向脂肪方向诱导分化,并通过油红O染色和吸光度值(OD值)检测其分化程度;qRT-PCR检测过氧化物酶体增殖物激活受体γ(PPARγ)、脂肪酸结合蛋白(FABP4)、CCAAT增强子结合蛋白(C/EBPα)mRNA表达水平;通过诱导的脂肪细胞与THP-1细胞共培养,加入1μg/ml的阿糖胞苷,在48 h测定其对肿瘤细胞的化疗抵抗作用。结果:通过油红O染色和测定吸光度值发现,3T3-L1细胞组脂质聚集量多于MSC组,且OD值大于MSC组(P0.05);qRTPCR结果显示,3T3-L1来源的脂肪细胞的成脂基因PPARγ、FABP4和C/EBPαmRNA的相对表达量高于人骨髓MSC的脂肪细胞(P0.05);共培养实验结果表明,含脂肪细胞组THP-1细胞存活数较对照组多(P0.05),且3T3-L1细胞组与MSC组之间具有统计学差异(P0.05)。结论:3T3-L1细胞的体外成脂能力比人骨髓MSC强;脂肪细胞具有促使THP-1细胞耐受化疗的作用,且在3T3-L1细胞组的作用大于人骨髓MSC组。
[Abstract]:Objective: to investigate the difference between 3T3-L1 cells and human bone marrow mesenchymal stem cells (MSCs) in adipogenesis and adipocytes in vitro. Methods: human bone marrow MSCs were isolated and purified by density gradient centrifugation and adherent culture, and the morphology of MSCs was observed under inverted microscope. Differentiation was induced to fat in vitro. The differentiation degree was detected by oil red O staining and OD value. The expression of peroxisome proliferator activated receptor (PPAR 纬) and fatty acid binding protein (FABP4) CCAAT enhancer protein (C / EBP 伪) were detected by qRT-PCR. After co-culture of adipocytes and THP-1 cells, 1 渭 g / ml cytarabine was added. Results: oil red O staining and absorbance measurement showed that the accumulation of lipid in the T3-L1 cell group was higher than that in the MSC group. The OD value was higher than that in MSC group. The results of RT-PCR showed that the relative expression levels of PPAR 纬 -FABP4 and C / EBP 伪 mRNA in adipocytes derived from 3T3-L1 were higher than those in human bone marrow MSC, and the results of co-culture showed that the relative expression of FABP4 and C / EBP 伪 mRNA in adipocytes derived from human bone marrow MSC was higher than that in human bone marrow MSC. The survival number of THP-1 cells in adipocyte group was more than that in control group, and there was statistical difference between 3T3-L1 cell group and MSC group. Conclusion the fat-forming ability of THP-1 cells in vitro is stronger than that of human bone marrow MSC, and adipocytes can promote the tolerance of THP-1 cells to chemotherapy. The effect of 3 T 3 L 1 cell group was greater than that of human bone marrow MSC group.
【作者单位】：苏州大学医学部;上海交通大学附属第六人民医院血液科;
【基金】：国家自然科学基金面上项目(81170507) 科技部重大仪器专项子课题(2013YQ03065109) 上海市科委项目(14142200702);上海市科委项目子课题(13DZ2293702)
【分类号】：R329.28

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