鸡卵清提取液中大于3ku蛋白促细胞升高表达多能基因Oct-3/4和Nanog

More than 3 ku proteins in chicken egg extract up-regulate expression of pluripotent genes Oct-3/4 and Nanog

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：Reprogramming somatic cells to generate pluripotent stem cells has a wide application in biomedical research. OBJECTIVE：To analyze the effect of different molecular weight proteins in chicken egg-white extract to elevate expression of pluripotent genes Oct-3/4 and Nanog in 293T cells. METHODS：The extracts of chicken egg-white were separated into more than 3 ku and less than 3 ku ingredients to be used for co-culture with 293T cells. There were four groups, 1×10^5 293T cells per wel , total 500μL. In the control group, 500μL culture medium was added;in the other three groups, 500μL chicken egg-white extract, more than 3 ku and less than 3 ku ingredients were respectively added. Quantitative PCR was used to determine the relative expression levels of pluripotent genes Nanog and Oct-3/4 in 293T cells. RESULTS AND CONCLUSION：By using co-culture method, more than 3 ku ingredients have a role to increase the expression of pluripotent genes Oct-3/4 and Nanog, but less than 3 ku ingredients cannot elevate the expression of pluripotent genes. This indicates that the ingredient of chicken egg-white extract to elevate the expression of pluripotent genes is more than 3 ku proteins.