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背景:将体细胞重编程为多能干细胞在生物医学研究领域有广泛的应用价值。目的:分析鸡卵清提取液中不同分子质量的蛋白促进293T细胞升高表达多能基因Oct-3/4和Nanog的作用。方法:分离鸡卵清提取液中大于3 ku和小于3 ku的成分,用于293T细胞共培养。实验分为4组:每孔加入1×105个293T细胞,总体积500μL。对照组加入500μL培养基;另外3个孔分别加500μL鸡卵清提取液、鸡卵清大于3 ku的成分和小于3 ku的成分。采用定量PCR检测多能基因Nanog和Oct-3/4的相对表达量。结果与结论:用共培养法大于3 ku的成分有促细胞升高表达多能基因Oct-3/4和Nanog的作用,但小于3 ku的成分没有促细胞升高表达多能基因的作用。提示在鸡卵清提取液中促细胞升高表达多能基因的成分是大于3 ku的蛋白质。
Background: The reprogramming of somatic cells into pluripotent stem cells has a wide range of applications in the field of biomedical research. OBJECTIVE: To analyze the effect of different molecular weight proteins in chicken egg white extract on the up-regulation of pluripotency gene Oct-3/4 and Nanog in 293T cells. Methods: Components of chicken egg white extract more than 3 ku and less than 3 ku were isolated and used for 293T cell co-culture. The experiment was divided into 4 groups: 1 × 105 293T cells were added into each well, the total volume was 500μL. The control group was supplemented with 500μL medium; the other three wells were added 500μL chicken egg white extract, chicken egg more than 3 ku ingredients and less than 3 ku ingredients. The relative expression levels of pluripotency gene Nanog and Oct-3/4 were detected by quantitative PCR. RESULTS AND CONCLUSION: Components co-cultured with more than 3 ku could promote the expression of pluripotency genes Oct-3/4 and Nanog, but less than 3 ku did not promote the expression of pluripotency gene. Prompted in chicken egg white extract cells to promote expression of pluripotency gene composition is greater than 3 ku protein.