Totipotent cells have more robust developmental potency than any other cell types, giving rise to both embryonic and extraembryonic tissues. Stable totipotent cell cultures and deciphering the principles of totipotency regulation would be invaluable to understand cell plasticity and lineage segregation in early development. Our approach of remodeling the pericentromeric heterochromatin and re-establishing the totipotency-specific broad H3K4me3 domains promotes the pluri-to-totipotency transition. Our protocol establishes a closer match of mouse 2-cell (2C) embryos than any other 2C-like cells. These totipotent-like stem cells (TLSCs) are stable in culture and possess unique molecular features of the mouse 2C embryo. Functionally, TLSCs are competent for germline transmission and give rise to both embryonic and extraembryonic lineages at high frequency. Therefore, TLSCs represent a highly valuable cell type for studies of totipotency and embryology.

全能细胞比任何其他细胞类型具有更强的发育潜能，从而产生胚胎和胚胎外组织。稳定的全能细胞培养和破译全能性调控的原理对于了解早期发育中的细胞可塑性和谱系分离非常宝贵。我们重塑着丝粒周围异染色质和重建全能特异性宽 H3K4me3 结构域的方法促进了多能性向全能性的转变。我们的协议建立了比任何其他 2C 样细胞更紧密的小鼠 2 细胞 (2C) 胚胎匹配。这些全能样干细胞 (TLSCs) 在培养中是稳定的，并具有小鼠 2C 胚胎的独特分子特征。在功能上，TLSCs 有能力进行种系传播，并以高频率产生胚胎和胚胎外谱系。因此，TLSCs 代表了一种非常有价值的细胞类型，用于研究全能性和胚胎学。