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Conversion between porcine naïve-like and primed ESCs and specific pluripotency marker identification.
In Vitro Cellular & Developmental Biology - Animal ( IF 1.5 ) Pub Date : 2020-05-19 , DOI: 10.1007/s11626-020-00448-3
Qiaoyu Chen 1, 2 , Hong Zhang 1, 2 , Haibin Jiang 1, 2 , Manling Zhang 1, 2, 3 , Junzheng Wang 1, 2 , Lihua Zhao 1, 2, 3 , Chenyu Wang 1, 2 , Manling Liu 1, 2 , Rongfeng Li 1, 2, 3
Affiliation  

Researchers currently lack standardized porcine-specific markers that would aid in distinguishing the naïve and primed states of porcine embryonic stem cells (ESCs). Here, we converted naïve-like porcine ESCs (nESCs, established in our lab) into primed-state cells, and we proposed a set of molecular criteria for evaluating the naïve porcine ESCs by comparing the two cell states. The reverse-primed porcine ESCs (rpESCs) are phenotypically stable and karyotypically intact. Alkaline phosphatase positivity and the ability to form embryonic bodies suggest that rpESCs still retain the capacity for self-renewal. Lineage-associated genes, such as Cdx2, Sox17, Eomes, Foxa, Fgf5, and Pitx2, exhibited significant expression in rpESCs. Nonetheless, LIF/3i-grown porcine ESCs treated with the small molecular weight inhibitors CHIR99021, PD0325901, and SB431542 expressed the greatest number of pluripotency marker genes, including Oct4, Sox2, Nog, Dppa5, Nr0b1, and Klf4, and at higher levels than were observed in rpESCs. Despite their general trend toward higher expression of critical pluripotency factors, the nESCs showed downregulation of Tbx3, Nanog, and c-Myc, which are considered typical naïve factors in other species. Entry of the nESCs into the developmentally primed state was also associated with a marked reduction in Lin28 expression. These findings extend the knowledge of porcine pluripotency markers and provide a backdrop for future analysis of naïve porcine pluripotency.

中文翻译:

猪幼稚样和引发的ESC之间的转换以及特定的多能性标记物鉴定。

目前,研究人员缺乏标准化的猪特异性标志物,这些标志物有助于区分猪胚胎干细胞(ESC)的原始状态和启动状态。在这里,我们将幼稚型猪胚胎干细胞(nESCs,在我们的实验室中建立)转化为初免状态细胞,并提出了一套分子标准,通过比较两种细胞状态来评估幼稚型猪胚胎干细胞。反向启动的猪胚胎干细胞(rpESCs)在表型上是稳定的,在核型上是完整的。碱性磷酸酶阳性和形成胚体的能力表明rpESCs仍然保留自我更新的能力。谱系相关基因,例如Cdx2,Sox17,Eomes,Foxa,Fgf5和Pitx2,在rpESCs中表现出显着表达。尽管如此,用小分子量抑制剂CHIR99021,PD0325901处理的LIF / 3i生长的猪ESC SB431542和SB431542表达的多能性标记基因数量最多,包括Oct4,Sox2,Nog,Dppa5,Nr0b1和Klf4,并且水平高于rpESCs中的水平。尽管它们普遍具有提高关键多能性因子表达的总体趋势,但nESCs仍下调了Tbx3,Nanog和c-Myc,它们被认为是其他物种中的典型幼稚因子。nESC进入发育启动状态也与Lin28表达的显着降低有关。这些发现扩展了猪多能性标志物的知识,并为未来的纯猪多能性分析提供了背景。尽管它们普遍具有提高关键多能性因子表达的总体趋势,但nESCs仍下调了Tbx3,Nanog和c-Myc,它们被认为是其他物种中的典型幼稚因子。nESC进入发育启动状态也与Lin28表达的显着降低有关。这些发现扩展了猪多能性标志物的知识,并为未来的纯猪多能性分析提供了背景。尽管它们普遍具有提高关键多能性因子表达的总体趋势,但nESCs仍下调了Tbx3,Nanog和c-Myc,它们被认为是其他物种中的典型幼稚因子。nESC进入发育启动状态也与Lin28表达的显着降低有关。这些发现扩展了猪多能性标志物的知识,并为未来的纯猪多能性分析提供了背景。
更新日期:2020-05-19
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