期刊
NATURE PROTOCOLS
卷 8, 期 6, 页码 1028-1041出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2013.049
关键词
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资金
- Human Frontier Science Program, US National Institutes of Health (NIH) [RO1-HD052115, RO1-DK084391]
- New York State Department of Health NYSTEM IDEA [C024318]
- March of Dimes Foundation [1-FY11-436]
- Medical Research Council (MRC)
- Centre for Trophoblast Research
- MRC [MC_UP_1202/9] Funding Source: UKRI
- Medical Research Council [MC_UP_1202/9] Funding Source: researchfish
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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