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Effects of cigarette smoke condensate on proliferation and pluripotency gene expression in mouse embryonic stem cells

机译:香烟烟雾凝结对小鼠胚胎干细胞增殖和多能基因表达的影响

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Abstract Background Embryonic stem cells (ESCs) are derived from the inner cell mass (ICM) of blastocysts. They can be used as valuable experimental models to test the effects of drugs, chemicals, and environmental contaminants such as cigarette smoke condensate (CSC) on preimplantation embryo development. The aim of this study was to evaluate the effect of CSC on ESCs derived from mice with different genetic backgrounds and maternal ages. Methods The study groups consisted of mouse?ESCs (mESCs) obtained from three sources: blastocysts developed from fertilized oocytes of two‐month‐old (2‐C57) and six‐month‐old (6‐C57) C57BL/6 inbred mice and those developed from fertilized oocytes of two‐month‐old (2‐NMRI) NMRI outbred mice. The groups of mESCs were exposed to 0.04, 4, and 40?μg/mL CSC. After exposure, we measured cell viability by 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl tetrazolium bromide assay and real‐time polymerase chain reaction for changes in expressions of Oct4, Sox2, Nanog, Ahr, Bax, Bcl2, TFAM , and POLG . The cell doubling time (DT) of these populations was also determined. Results We observed that CSC changed proliferation and DT in the 2‐C57 and 6‐C57 cells. There was no change in 2‐NMRI cells. Exposure to CSC caused changes in the gene expressions and induced apoptosis in all three cell lines. Conclusion Based on the results of the study, it can be concluded that CSC has an effect on the viability, DT and gene expression patterns in mouse ESCs and its effects vary based on the genetic background and maternal age of isolated mouse ESCs.
机译:摘要背景胚胎干细胞(ESC)来自胚泡的内部电池质量(ICM)。它们可用作有价值的实验模型来测试药物,化学品和环境污染物如香烟烟雾缩合物(CSC)的效果对胚胎发育。本研究的目的是评估CSC对来自不同遗传背景和母体年龄的小鼠的ESC的作用。方法研究组由小鼠(MESCS)组成,从三个来源获得:从两月大(2-C57)和六个月大(6-C57)C57BL / 6次闭合小鼠的受精卵母细胞产生的胚泡从两月大(2-NMRI)NMRI分区老鼠的受精卵母细胞产生的那些。将麦塞斯组暴露于0.04,4和40Ωμg/ ml CSC。曝光后,我们通过3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四唑溴化沸石测定和实时聚合酶链反应测量细胞活力,以及Oct4,Sox2,Nanog,AHR, bax,bcl2,tfam和polg。还确定了这些群体的细胞倍增时间(DT)。结果我们观察到CSC改变了2-C57和6-C57细胞中的增殖和DT。 2-NMRI细胞没有变化。接触CSC导致基因表达的变化并在所有三种细胞系中诱导细胞凋亡。结论基于该研究的结果,可以得出结论,CSC对小鼠ESC中的活力,DT和基因表达模式的影响及其效果基于孤立的鼠标ESC的遗传背景和母体年龄而变化。

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