Guo Xin, Zhang Yan-min, Qi Zheng-yu, Qin Jie, Cui Guang-hui, Gui Yao-ting, Cai Zhi-ming
Key Laboratory of Male Reproduction and Genetics of Guangdong Province, Shenzhen Hospital of Peking University, Shenzhen, Guangdong 518036, China.
Zhonghua Nan Ke Xue. 2011 Nov;17(11):966-72.
To investigate whether mouse-induced pluripotent stem (iPS) cell line IP14D-1 has the potential to differentiate into induced primordial germ cells (iPGCs), and to explore the changes in the expression of iPGCs-differentiation associated genes and their possible mechanisms.
Undifferentiated IP14D-1 was cultured to proliferate and then differentiated to form 4-, 7- and 9-day-old induced embryoid bodies (iEBs) in vitro, respectively. RT-PCR and immunofluorescence were used to detect the expressions of Lin28, Blimpl, Stra8 and Mvh, as well as the localization of the corresponding protein in iEBs.
The expression of Blimpl was higher than that of Lin28 in the undifferentiated IP14D-1 and mouse embryonic stem cells (mESCs). Mvh and Stra8 as well as mESCs and EBs were also expressed in IP14D-1 and iEBs, but with no significant differences. The expression of Lin28 was gradually increased in the IP14D-1-derived iEBs from 4 to 7 days, but decreased at 9 days, and the expression of Blimp1 was gradually reduced with the prolonged growing time of iEBs.
A stable system was established for the culture and differentiation of IP14D-1 and IP14D-1-derived iEBs. The expressions of Lin28, Blimp1, Mvh and Stra8 were not significantly different between the undifferentiated IP14D-1 and mESCs, nor were the expressions of Mvh and Stra8 between iEBs and EBs. IP14D-1 and iEBs had the potential to differentiate into iPGCs, which increased in number in the 7-day-old iEBs, and the expression of iPGC-differentiation associated Lin28 became lower in the older iEBs.
研究小鼠诱导多能干细胞系IP14D-1是否具有分化为诱导原始生殖细胞(iPGCs)的潜力,并探讨iPGCs分化相关基因表达的变化及其可能机制。
将未分化的IP14D-1进行培养增殖,然后分别在体外分化形成4、7和9日龄的诱导胚状体(iEBs)。采用RT-PCR和免疫荧光检测iEBs中Lin28、Blimpl、Stra8和Mvh的表达以及相应蛋白的定位。
在未分化的IP14D-1和小鼠胚胎干细胞(mESCs)中,Blimpl的表达高于Lin28。Mvh和Stra8以及mESCs和EBs在IP14D-1和iEBs中也有表达,但无显著差异。在IP14D-1来源的iEBs中,Lin28的表达在4至7天逐渐升高,但在9天时降低,而Blimp1的表达随着iEBs生长时间的延长逐渐降低。
建立了IP14D-1及其来源的iEBs培养和分化的稳定体系。未分化的IP14D-1与mESCs之间Lin28、Blimpl、Mvh和Stra8的表达无显著差异,iEBs与EBs之间Mvh和Stra8的表达也无显著差异。IP14D-1和iEBs具有分化为iPGCs的潜力,在7日龄的iEBs中数量增加,且随着iEBs年龄增长,iPGC分化相关的Lin28表达降低。