维甲酸增加人胚胎干细胞形成拟胚体中生殖细胞特异基因标记

Retinoic acid increases expression of germ cell-specific markers of embryoid body differentiated from human embryonic stem cells

目的探讨维甲酸在增加人胚胎干细胞(hESCs)形成拟胚体中生殖细胞特异基因标记中的作用。方法 hESCs在含有维甲酸或无维甲酸的分化液中悬浮培养形成拟胚体,采用实时荧光定量PCR技术检测未分化因子OCT-4及生殖细胞特异性标记VASA、SCP3、GDF9和TEKT1的表达情况。结果在拟胚体形成过程中,未分化因子OCT-4表达下降,但生殖母细胞标记VASA、减数分裂标记SCP3及减数分裂后标记GDF9和TEKT1表达均增加。在含有维甲酸的分化系统中,生殖细胞特异性标记表达均增加。培养5d后,生殖细胞特异性标记VASA、SCP3、GDF9和TEKT1分别增加9.3、6.9、7.2和11.8倍。结论 hESCs具有分化为原始生殖细胞(PGCs)和早期配子的能力。维甲酸能增加hESCs形成拟胚体中生殖细胞特异性标记的表达。

Objective： To study the effect of retinoic acid （RA） on the embryoid body differentiated from human embryonic stem cells Methods： hESCs were differentiated as embryoid bodies （atRA） or without atRA. Then the expressions of OCT4, evaluated by RT-PCR. expression of germ cell-specific markers ot （hESCs）. （EBs） in suspension VASA, SCP3, GDF9 with all-trans RA and TEKT1 were Results： The expressions of germ cell-specific markers including the gonocyte marker VASA, the meiotic marker SCP3, the postmeiotic markers GDF9 and TEKT1 were all increased in the presence and absence of RA during EB differehtiation was progressed. In addition, the expressions of these markers were increased averages of 9.3, 6.9, 7.2 and 11.8 fold respectively in the presence of RA compared with the absence of RA during 5 days differentiation. Conclusions： The results indicated that hESCs might have the potential to differentiate to PGCs and early gametes. And RA could increase the expression of germ cell-specific markers of embryoid body differentiated from hESCs.