Conversion between porcine naive-like and primed ESCs and specific pluripotency marker identification
Conversion between porcine naive-like and primed ESCs and specific pluripotency marker identification
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DOI：
10.1007/s11626-020-00448-3
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发表时间：
2020-05-19
影响因子：
2.1
通讯作者：
Li，Rongfeng
中科院分区：
文献类型：
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作者：
Chen，Qiaoyu;Zhang，Hong;Li，Rongfeng
研究人员目前缺乏标准化的猪特异性标记物，可以帮助区分猪胚胎干细胞（ESC）的幼稚和启动状态。在这里，我们将幼稚样猪胚胎干细胞（nESC，在我们的实验室建立）转化为致敏状态的细胞，我们提出了一套分子标准，通过比较两种细胞状态来评估幼稚猪胚胎干细胞。反向致敏的猪ESC（rpESC）是表型稳定且核型完整的。碱性磷酸酶阳性和形成胚胎体的能力表明，rpESCs仍然保留自我更新的能力。谱系相关基因如Cdx 2、Sox 17、Eomes、Foxa、Fgf 5和Pitx 2在rpESCs中表现出显著的表达。尽管如此，用小分子量抑制剂CHIR 99021、PD 0325901和SB 431542处理的LIF/3 i生长的猪ESC表达了最多数量的多能性标记基因，包括Oct 4、Sox 2、Nog、Dppa 5、Nr 0 b1和Klf 4，并且表达水平高于在rpESC中观察到的水平。尽管nESC的关键多能性因子的表达总体上呈较高趋势，但它们显示出Tbx 3、Nanog和c-Myc的下调，这些因子被认为是其他物种中典型的幼稚因子。nESC进入发育启动状态也与Lin 28表达的显著减少有关。这些发现扩展了猪多能性标记的知识，并为未来分析幼稚猪多能性提供了背景。
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 asCdx2,Sox17,Eomes,Foxa,Fgf5, andPitx2, 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, includingOct4,Sox2,Nog,Dppa5,Nr0b1, andKlf4, and at higher levels than were observed in rpESCs. Despite their general trend toward higher expression of critical pluripotency factors, the nESCs showed downregulation ofTbx3,Nanog, andc-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 inLin28expression. These findings extend the knowledge of porcine pluripotency markers and provide a backdrop for future analysis of naïve porcine pluripotency.