Southern Medical University Central Laboratory, Southern Medical University, Guangzhou, China.
School of Basic Medical Science, Southern Medical University, Guangzhou, China.
Cytometry A. 2022 Aug;101(8):682-691. doi: 10.1002/cyto.a.24551. Epub 2022 Mar 25.
Mouse embryonic stem cells (ESCs) and epiblast stem cells (EpiSCs) are both pluripotent stem cells from early embryos. Another type of pluripotent stem cells, which are similar with EpiSCs and derive from pre-implantation embryos in feeder-free and chemically defined medium containing Activin A and basic fibroblast growth factors (bFGF), is termed as AFSCs. The pluripotency and self-renewal maintenance of ESCs rely on Leukemia inhibitory factor (LIF)/STAT/BMP4/SMAD signaling, while the pluripotency and self-renewal maintenance of EpiSCs and AFSCs rely on bFGF and Activin/Nodal signaling. However, the establishment efficiency of AFSCs lines is low. In this study, we stimulated early embryos by 2i/LIF (CHIR99021 + PD0325901 + LIF) and Activin A + bFGF respectively, to change the cell fate in inner cell mass (ICM). The "fate changed embryos" by 2i/LIF can efficiently produce AFSCs in feeder-free and chemically defined medium, but the efficiency of embryos treated with Activin A + bFGF were poor. The AFSCs from fate-changed embryos share similar molecular characteristics with conventional AFSCs and EpiSCs. Our results suggest that the advanced stimulation of 2i/LIF and the premature stimulation of Activin A + bFGF contribute to capturing the pluripotent stem cells in early embryos, and the FGF/MAPK signaling dominate early embryo development. Our study provides a new approach to capturing pluripotency from pre-implantation embryos.
小鼠胚胎干细胞（ESCs）和胚胎外胚层干细胞（EpiSCs）均来自早期胚胎的多能干细胞。另一种多能干细胞，与 EpiSCs 相似，源自无饲养层和含有激活素 A 和碱性成纤维细胞生长因子（bFGF）的化学定义培养基中的着床前胚胎，被称为 AFSCs。ESCs 的多能性和自我更新维持依赖于白血病抑制因子（LIF）/STAT/BMP4/SMAD 信号通路，而 EpiSCs 和 AFSCs 的多能性和自我更新维持依赖于 bFGF 和激活素/Nodal 信号通路。然而，AFSCs 系的建立效率很低。在这项研究中，我们分别用 2i/LIF（CHIR99021+PD0325901+LIF）和激活素 A+bFGF 刺激早期胚胎，以改变内细胞团（ICM）中的细胞命运。2i/LIF 处理的“命运改变胚胎”可以在无饲养层和化学定义培养基中有效地产生 AFSCs，而用激活素 A+bFGF 处理的胚胎效率则较差。命运改变胚胎来源的 AFSCs 与传统的 AFSCs 和 EpiSCs 具有相似的分子特征。我们的结果表明，2i/LIF 的高级刺激和激活素 A+bFGF 的过早刺激有助于从早期胚胎中捕获多能干细胞，并且 FGF/MAPK 信号通路主导早期胚胎发育。我们的研究为从着床前胚胎中捕获多能性提供了一种新方法。
