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本研究采用超高速离心法从人胚胎干细胞培养液上清分离提纯出外泌体,应用透射电镜、免疫印记和RT-PCR方法检测其生物学特性。结果表明,在透射电镜下观察到人胚胎干细胞外泌体呈茶盏状,大小不均匀,直径30～100 nm,有完整的膜结构,内含低密度物质;人胚胎干细胞外泌体表达外泌体标志抗原CD63、TSG101和多能性转录因子OCT4、SOX2、NANOG、KLF4;干细胞外泌体呈剂量性依赖方式促进小鼠胚胎成纤维细胞增殖。本研究为进一步研究人胚胎干细胞外泌体的生物学特征和功能提供有价值的生物学信息。
Abstract：Exosomes were isolated from cell culture meidium by ultracentrifugation, and the biological characteristics of exosomes were identified by transmission electron microscopy, Western blotting and RT-PCR. The results showed that human embryonic stem cell derived exosome displayed tipically tea-cup shape, with a diameter of about 30～100 nm, and posessed complete membrane with low density content under transmission elctron microscopy. Human embryonic stem cell derived exosome experessed mark proteins(CD63, TSG101), and pluripotent factors(OCT4, SOX2, NANOG and KLF4). Meanwhile, human embryonic stem cell derived exosome promoted the proliferation of mouse embrynic fibroblasts in a does-dependent manner. This study provided valuable information for the further study of biological characteristic and functions of human embryonic stem cell derived exosome.
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基本信息:
DOI：10.13417/j.gab.039.005286
中图分类号:R329.2
引用信息:
[1]农微,吴剑婷,吴青萍,等.人胚胎干细胞外泌体的分离与生物学功能鉴定[J].基因组学与应用生物学,2020,39(11):5286-5290.DOI:10.13417/j.gab.039.005286.
基金信息:
广西高校中青年教师基础能力提升项目(2017KY0285);; 广西中医药大学青年基金项目(2017QN017);; 广西大学大学生创新计划(201710593078)共同资助
2020-11-25
2020-11-25