Xuemei Liu, Jing Yue, Bei Xu, Juan Hu, Xinling Ren, Qun Liu, Guijin Zhu
Reproductive Medicine Center, Yantai Yuhuangding Hospital, Yantai 264000, China.
Reproductive Medicine Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Iran J Reprod Med. 2013 Nov;11(11):905-12.
Creation of artificial gametes may provide a universal solution for these patients with no gametes. Stem cell technology may provide a way to obtain fully functional gametes. Retinoic acid (RA) can initiate meiosis. Several studies have demonstrated that RA can promote sperm cells differentiation from mouse embryonic stem cells (mESCs) and other cells from human embryonic stem cells (hESCs).
We sought to determine whether RA could promote differentiation of germ cells from hESCs.
hESCs were differentiated as embryoid bodies (EBs) in suspension with all-trans RA (atRA) or without atRA for 0, 1, 3, 5 and 7 days, and then the expression of VASA, SCP3, GDF9 and TEKT1 were compared by real-time PCR. The statistical differences were evaluated by one way ANOVA.
The expression of germ cell-specific markers including the gonocyte marker VASA, the meiotic marker SCP3, and post meiotic markers, GDF9 and TEKT1, all increased in the presence and absence of RA as EB differentiation progressed. In addition, the expression of these markers increased an average of 9.3, 6.9, 7.2 and 11.8 fold respectively in the presence of RA, compared to the absence of RA, over 5 days differentiation.
Our results indicate that hESCs may have the potential to differentiate to primordial germ cells (PGCs) and early gametes. RA can improve germ cells differentiation from hESCs.
人工配子的产生可能为这些没有配子的患者提供一个通用的解决方案。干细胞技术可能提供一种获得功能完全正常的配子的方法。视黄酸(RA)可以启动减数分裂。多项研究表明,RA可以促进小鼠胚胎干细胞(mESCs)分化为精子细胞,以及人胚胎干细胞(hESCs)分化为其他细胞。
我们试图确定RA是否能促进hESCs向生殖细胞分化。
将hESCs在含有或不含有全反式视黄酸(atRA)的情况下悬浮分化为胚状体(EBs),分别培养0、1、3、5和7天,然后通过实时PCR比较VASA、SCP3、GDF9和TEKT1的表达。采用单因素方差分析评估统计学差异。
随着EB分化的进行,包括生殖母细胞标志物VASA、减数分裂标志物SCP3以及减数分裂后标志物GDF9和TEKT1在内的生殖细胞特异性标志物的表达,无论有无RA均增加。此外,在5天的分化过程中,与无RA相比,有RA时这些标志物的表达分别平均增加了9.3、6.9、7.2和11.8倍。
我们的结果表明,hESCs可能具有分化为原始生殖细胞(PGCs)和早期配子的潜力。RA可以促进hESCs向生殖细胞分化。