Molecular and embryology studies have demonstrated that mouse pre-implantation embryo development is a process of progressive cell fate determination. At the time of implantation, three cell lineages are present in the developing blastocyst: the trophectoderm (TE), the epiblast (Epi) and the primitive endoderm (PrE). From these early embryo cells, trophoblast stem (TS) cells, embryonic stem (ES) cells and extra-embryonic endoderm stem (XEN) cells can be derived. Recently, we derived stem cells with blastomere-like features from mouse cleavage-stage embryos, which we named expanded-potential stem cells (EPSCs). Here, we provide detailed protocols that describe how to establish EPSCs from single eight-cell-stage blastomeres or whole eight-cell pre-implantation mouse embryos, or by conversion of mouse ES cells or induced pluripotent stem (iPS) cells reprogrammed from fibroblasts. It takes 2-3 weeks to derive EPSCs from each cell source. The EPSCs derived from these protocols can differentiate into all embryonic and extra-embryonic lineages when implanted into chimeras. Furthermore, bona fide TS and XEN cell lines can be derived from EPSCs in vitro.

中文翻译：

---

从小鼠植入前的胚胎或胚胎干细胞体外建立扩增潜能干细胞。

分子和胚胎学研究表明，小鼠植入前胚胎的发育是决定细胞命运的一个过程。在植入时，发育中的胚泡中存在三种细胞谱系：滋养外胚层（TE），表皮细胞（Epi）和原始内胚层（PrE）。从这些早期胚胎细胞中，可以得到滋养层干（TS）细胞，胚胎干（ES）细胞和胚外内胚层干（XEN）细胞。最近，我们从小鼠卵裂期的胚胎中获得了具有卵裂球样特征的干细胞，我们将其命名为扩展电位干细胞（EPSC）。在这里，我们提供详细的协议，描述如何从单个八细胞阶段的卵裂球或整个八细胞植入前的小鼠胚胎中建立EPSC，或通过转化成纤维细胞重新编程的小鼠ES细胞或诱导多能干（iPS）细胞进行转化。从每个细胞来源中提取EPSC需要2-3周的时间。这些协议衍生的EPSC植入嵌合体后可以分化为所有胚胎和胚外谱系。此外，真正的TS和XEN细胞系可以在体外从EPSC获得。