Effects of sustained hypoxia on proliferation of mouse embryonic fibroblasts and preparation of feeder layers

  title={Effects of sustained hypoxia on proliferation of mouse embryonic fibroblasts and preparation of feeder layers},
  author={Han‐qing Wei and Yijin Pei and Dandan Wang and Yang Jiang and Chun Wang and Hong‐mei Li},
  journal={Journal of Clinical Rehabilitative Tissue Engineering Research},
摘要 背景:在传统的胚胎干细胞培养体系中,饲养层细胞制备和胚胎干细胞的培养扩增大都是在常氧条件下进行 的,氧培养条件的改变有可能影响饲养层细胞,从而改变胚胎干细胞的生长特性或分化能力,但尚未见到这 方面的系统研究报道。 目的:观察低氧培养对饲养层细胞及小鼠胚胎干细胞干性维持的影响。 方法:原代小鼠胚胎成纤维细胞分为常氧组(体积分数 20%O2)和低氧组(体积分数 5%O2)持续传代培养,绘制生 长曲线,检测活性氧水平和线粒体膜电位。将小鼠胚胎干细胞分为常氧组(饲养层为常氧组小鼠胚胎成纤维细胞, 体积分数 20%O2条件下培养)和低氧组(饲养层为低氧组小鼠胚胎成纤维细胞,体积分数 5%O2条件下培养),观 察胚胎干细胞的生长形态,检测干细胞多能性指标 Oct4、Sox2 以及低氧诱导因子 HIF-1α mRNA 表达。 结果与结论:1与常氧组相比,低氧组小鼠胚胎成纤维细胞增殖较快,线粒体膜电位上升,产生活性氧减少 (P < 0.05);2胚胎干细胞碱性磷酸酶染色阳性,Oct4、Sox2 高表达,低氧组形成的中小集落及 HIF-1α mRNA 表达比常氧组显著增多(P < 0.05);3… Expand

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