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Wnt pathway modulation generates blastomere-derived mouse embryonic stem cells with different pluripotency features
Journal of Assisted Reproduction and Genetics ( IF 3.1 ) Pub Date : 2020-10-12 , DOI: 10.1007/s10815-020-01964-7
Marta Vila-Cejudo 1, 2 , Sandra Alonso-Alonso 1 , Anna Pujol 3 , Josep Santaló 1 , Elena Ibáñez 1
Affiliation  

Purpose

This study aimed to determine the role of Wnt pathway in mouse embryonic stem cell (mESC) derivation from single blastomeres isolated from eight-cell embryos and in the pluripotency features of the mESC established.

Methods

Wnt activator CHIR99021, Wnt inhibitor IWR-1-endo, and MEK inhibitor PD0325901 were used alone or in combination during ESC derivation and maintenance from single blastomeres biopsied from eight-cell embryos. Alkaline phosphatase activity, FGF5 levels, expression of key pluripotency genes, and chimera formation were assessed to determine the pluripotency state of the mESC lines.

Results

Derivation efficiencies were highest when combining pairs of inhibitors (15–24.7%) than when using single inhibitors or none (1.4–10.1%). Full naïve pluripotency was only achieved in CHIR- and 2i-treated mESC lines, whereas IWR and PD treatments or the absence of treatment resulted in co-existence of naïve-like and primed-like pluripotency features. IWR + CHIR- and IWR + PD-treated mESC displayed features of primed pluripotency, but IWR + CHIR-treated lines were able to generate germline-competent chimeric mice, resembling the predicted properties of formative pluripotency.

Conclusion

Wnt and MAPK pathways have a key role in the successful derivation and pluripotency features of mESC from single precompaction blastomeres. Modulation of these pathways results in mESC lines with various degrees of naïve-like and primed-like pluripotency features.



中文翻译:

Wnt 通路调节产生具有不同多能性特征的卵裂球来源的小鼠胚胎干细胞

目的

本研究旨在确定 Wnt 通路在从八细胞胚胎中分离出的单个卵裂球的小鼠胚胎干细胞 (mESC) 衍生中的作用,以及在建立的 mESC 的多能性特征中的作用。

方法

Wnt 激活剂 CHIR99021、Wnt 抑制剂 IWR-1-endo 和 MEK 抑制剂 PD0325901 在从八细胞胚胎活检的单个卵裂球进行 ESC 衍生和维持期间单独或联合使用。评估碱性磷酸酶活性、FGF5 水平、关键多能性基因的表达和嵌合体形成,以确定 mESC 系的多能性状态。

结果

与使用单一抑制剂或不使用抑制剂 (1.4–10.1%) 相比,组合抑制剂对 (15–24.7%) 的衍生效率最高。完全初始多能性仅在 CHIR 和 2i 处理的 mESC 系中实现,而 IWR 和 PD 处理或不进行治疗导致初始样和引发样多能性特征共存。IWR + CHIR- 和 IWR + PD 处理的 mESC 显示出引发多能性的特征,但 IWR + CHIR 处理的细胞系能够产生具有种系能力的嵌合小鼠,类似于形成性多能性的预测特性。

结论

Wnt 和 MAPK 通路在 mESC 从单个预压实卵裂球的成功衍生和多能性特征中起关键作用。这些通路的调节导致 mESC 系具有不同程度的幼稚样和致敏样多能性特征。

更新日期:2020-10-12
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