本文選題：胰島素樣生長(zhǎng)因子-1 + 誘導(dǎo)多潛能干細(xì)胞�。� 參考：《華中科技大學(xué)》2012年碩士論文

【摘要】：目的：研究胰島素樣生長(zhǎng)因子-1（IGF-1）在iPS細(xì)胞重編程過(guò)程中的作用。 方法：1.利用含IGF-1的成纖維細(xì)胞培養(yǎng)基培養(yǎng)小鼠成纖維細(xì)胞3天,此為實(shí)驗(yàn)組。 2.利用不含IGF-1成纖維細(xì)胞培養(yǎng)基在相同條件下培養(yǎng)小鼠成纖維細(xì)胞3天，此為對(duì)照組。 3.分別對(duì)兩組成纖維細(xì)胞用含Oct4，Sox2，Klf4和c-Myc因子的四種逆 轉(zhuǎn)錄病毒顆粒進(jìn)行感染，對(duì)形成的iPS細(xì)胞進(jìn)行鑒定。 結(jié)果：對(duì)照組感染病毒后第21天出現(xiàn)類似胚胎干(embryonic stem；ES)細(xì)胞形態(tài)的克隆，共計(jì)4個(gè)克隆出現(xiàn)，誘導(dǎo)效率約為0.01%。IGF-1處理后的實(shí)驗(yàn)組感染病毒后第18天出現(xiàn)類似ES細(xì)胞形態(tài)的克隆，共計(jì)15個(gè)克隆出現(xiàn)，誘導(dǎo)效率約為0.05%。實(shí)驗(yàn)組的AP染色及SSEA-1、Oct4染色鑒定均為陽(yáng)性，且誘導(dǎo)效率較對(duì)照組提高近5倍。 結(jié)論：IGF-1在iPS細(xì)胞重編程中發(fā)揮著重要作用，其可以提高四因子體系的誘導(dǎo)效率。 目的：研究胰島素樣生長(zhǎng)因子-1(IGF-1)在降低iPS細(xì)胞致癌性方面的作用。 方法：1.利用含IGF-1的成纖維細(xì)胞培養(yǎng)基培養(yǎng)成纖維細(xì)胞3天，此為實(shí)驗(yàn)組。 2.在相同條件下，用不含IGF-1的成纖維細(xì)胞培養(yǎng)基培養(yǎng)小鼠成纖維細(xì)胞 3天，此為對(duì)照組。 3.分別對(duì)兩組成纖維細(xì)胞用含Oct4，Sox2和Klf4因子的三種逆轉(zhuǎn)錄病毒顆粒進(jìn)行感染，對(duì)形成的iPS細(xì)胞進(jìn)行鑒定。 結(jié)果：對(duì)照組經(jīng)病毒感染后未出現(xiàn)具有ES細(xì)胞形態(tài)的克隆。IGF-1處理后的實(shí)驗(yàn)組感染病毒后第20天出現(xiàn)類似ES細(xì)胞形態(tài)的克隆，共計(jì)3個(gè)克隆出現(xiàn)，，誘導(dǎo)效率約為0.01%。實(shí)驗(yàn)組的iPS細(xì)胞AP染色及SSEA-1、Oct4染色鑒定均為陽(yáng)性。 結(jié)論：IGF-1在降低iPS細(xì)胞的致癌性方面起著重要作用，且聯(lián)合三因子誘導(dǎo)iPS細(xì)胞，重編程效率沒(méi)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.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R329.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 周琪;曾凡一;;通過(guò)四倍體補(bǔ)償實(shí)驗(yàn)證明iPS細(xì)胞具有發(fā)育全能性[J];中國(guó)基礎(chǔ)科學(xué);2010年03期

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