The Anne McLaren Laboratory for Regenerative Medicine, University of Cambridge, Cambridge, UK.
Nat Protoc. 2013 Jun;8(6):1028-41. doi: 10.1038/nprot.2013.049. Epub 2013 May 2.
At the time of implantation in the maternal uterus, the mouse blastocyst possesses an inner cell mass comprising two lineages: epiblast (Epi) and primitive endoderm (PrE). Representative stem cells derived from these two cell lineages can be expanded and maintained indefinitely in vitro as either embryonic stem (ES) or XEN cells, respectively. Here we describe protocols that can be used to establish XEN cell lines. These include the establishment of XEN cells from blastocyst-stage embryos in either standard embryonic or trophoblast stem (TS) cell culture conditions. We also describe protocols for establishing XEN cells directly from ES cells by either retinoic acid and activin-based conversion or by overexpression of the GATA transcription factor Gata6. XEN cells are a useful model of PrE cells, with which they share gene expression, differentiation potential and lineage restriction. The robust protocols for deriving XEN cells described here can be completed within 2-3 weeks.
在植入母体子宫时，小鼠囊胚具有内细胞团，包含两个谱系：上胚层（Epi）和原始内胚层（PrE）。分别从这两个细胞谱系衍生的代表性干细胞可以在体外无限期地扩增和维持，分别为胚胎干细胞（ES）或 XEN 细胞。在这里，我们描述了可以用来建立 XEN 细胞系的方案。这些方案包括在标准胚胎或滋养层干细胞（TS）培养条件下，从囊胚期胚胎中建立 XEN 细胞。我们还描述了通过视黄酸和激活素为基础的转化或通过过表达 GATA 转录因子 Gata6 直接从 ES 细胞建立 XEN 细胞的方案。XEN 细胞是原始内胚层细胞的有用模型，它们具有相似的基因表达、分化潜能和谱系限制。这里描述的用于衍生 XEN 细胞的强大方案可以在 2-3 周内完成。
