胰岛素样生长因子-1联合外源性转录因子诱导小鼠iPS细胞的研究

发布时间：2018-06-22 16:01

本文选题：胰岛素样生长因子-1 + 诱导多潜能干细胞　；参考：《华中科技大学》2012年硕士论文

【摘要】：目的：研究胰岛素样生长因子-1（IGF-1）在iPS细胞重编程过程中的作用。方法：1.利用含IGF-1的成纤维细胞培养基培养小鼠成纤维细胞3天,此为实验组。 2.利用不含IGF-1成纤维细胞培养基在相同条件下培养小鼠成纤维细胞3天，此为对照组。 3.分别对两组成纤维细胞用含Oct4，Sox2，Klf4和c-Myc因子的四种逆转录病毒颗粒进行感染，对形成的iPS细胞进行鉴定。结果：对照组感染病毒后第21天出现类似胚胎干(embryonic stem；ES)细胞形态的克隆，共计4个克隆出现，诱导效率约为0.01%。IGF-1处理后的实验组感染病毒后第18天出现类似ES细胞形态的克隆，共计15个克隆出现，诱导效率约为0.05%。实验组的AP染色及SSEA-1、Oct4染色鉴定均为阳性，且诱导效率较对照组提高近5倍。结论：IGF-1在iPS细胞重编程中发挥着重要作用，其可以提高四因子体系的诱导效率。目的：研究胰岛素样生长因子-1(IGF-1)在降低iPS细胞致癌性方面的作用。方法：1.利用含IGF-1的成纤维细胞培养基培养成纤维细胞3天，此为实验组。 2.在相同条件下，用不含IGF-1的成纤维细胞培养基培养小鼠成纤维细胞 3天，此为对照组。 3.分别对两组成纤维细胞用含Oct4，Sox2和Klf4因子的三种逆转录病毒颗粒进行感染，对形成的iPS细胞进行鉴定。结果：对照组经病毒感染后未出现具有ES细胞形态的克隆。IGF-1处理后的实验组感染病毒后第20天出现类似ES细胞形态的克隆，共计3个克隆出现，，诱导效率约为0.01%。实验组的iPS细胞AP染色及SSEA-1、Oct4染色鉴定均为阳性。结论：IGF-1在降低iPS细胞的致癌性方面起着重要作用，且联合三因子诱导iPS细胞，重编程效率没有降低。
[Abstract]:Aim: to study the role of insulin-like growth factor-1 (IGF-1) in the reprogramming of iPS cells. Method 1: 1. Mouse fibroblasts were cultured in IGF-1 medium for 3 days. Mouse fibroblasts were cultured in IGF-1 free fibroblast medium under the same conditions for 3 days. The two fibroblasts were infected with four reverse transcriptional virus particles containing Oct4N Sox2Klf4 and c-Myc factor, and the resulting iPS cells were identified. Results: the morphological clones of embryonic stem es cells appeared in the control group on the 21st day after infection. A total of 4 clones appeared. The induction efficiency was about 0.01. IGF-1 treated experimental group showed similar es cell morphology clone on the 18th day after infection. A total of 15 clones appeared and the induction efficiency was about 0.05%. AP staining and SSEA-1 Oct4 staining were positive in the experimental group, and the induction efficiency was 5 times higher than that in the control group. ConclusionTwo one IGF-1 plays an important role in the reprogramming of iPS cells, and it can improve the induction efficiency of the four-factor system. Aim: to study the effect of insulin-like growth factor-1 (IGF-1) on the carcinogenicity of iPS cells. Method 1: 1. The fibroblasts were cultured in IGF-1 medium for 3 days. Under the same conditions, mouse fibroblasts were cultured on fibroblast medium without IGF-1 for 3 days, which was the control group. The two fibroblasts were infected with three kinds of retrovirus particles containing Oct4G Sox2 and Klf4, and the resulting iPS cells were identified. Results: there were no clones with es cell morphology in the control group. The experimental group infected with IGF-1 had similar es cell morphology on the 20th day after infection. A total of 3 clones appeared, and the induction efficiency was about 0.01. AP staining and SSEA-1 Oct 4 staining of iPS cells were positive in the experimental group. Conclusion the weight IGF-1 plays an important role in decreasing the carcinogenicity of iPS cells, and the reprogramming efficiency of iPS cells induced by the combination of three factors is not decreased.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R329.2

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