Derivation of extraembryonic endoderm stem (XEN) cells from mouse embryos and embryonic stem cells.

Derivation of extraembryonic endoderm stem (XEN) cells from mouse embryos and embryonic stem cells.
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DOI:
10.1038/nprot.2013.049
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发表时间:
2013-06
期刊:
影响因子:
14.8
通讯作者:
Hadjantonakis, Anna-Katerina
Hadjantonakis, Anna-Katerina
中科院分区:
生物学1区
文献类型:
--
作者:
Niakan, Kathy K.;Schrode, Nadine;Cho, Lily T. Y.;Hadjantonakis, Anna-Katerina

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在母体子宫着床时,小鼠囊胚具有由两种细胞系组成的内细胞群:外胚层(Epi)和原始内胚层(PrE)。来自这两种细胞系的代表性干细胞可以分别作为胚胎干细胞(ES)或XEN细胞在体外无限扩增和维持。在这里,我们描述了可用于建立XEN细胞系的协议。这些方法包括在标准胚胎或滋养细胞干细胞(TS)细胞培养条件下从囊胚期胚胎培养XEN细胞。我们还描述了通过视黄酸和激活素转化或通过过表达GATA转录因子Gata6直接从ES细胞建立XEN细胞的方案。XEN细胞是PrE细胞的一个有用的模型,它们具有相同的基因表达、分化潜力和谱系限制。本文所述的可靠的XEN细胞提取方案可在2-3周内完成。
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.
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