title: Derivation of extraembryonic endoderm stem (XEN) cells from mouse embryos and embryonic stem cells.
abstract: 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.
authors: The Anne McLaren Laboratory for Regenerative Medicine, University of Cambridge, Cambridge, UK. kniakan@nimr.mrc.ac.uk
authors: Kathy K
authors: KK
authors: Niakan
authors: Nadine
authors: N
authors: Schrode
authors: Lily T Y
authors: LT
authors: Cho
authors: Anna-Katerina
authors: AK
authors: Hadjantonakis
mesh_terms: D028341:Activins / Q000378:metabolism; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation / Q000379:methods*; D004622:Embryo, Mammalian / Q000166:cytology*; D053595:Embryonic Stem Cells / Q000166:cytology*; D004707:Endoderm / Q000166:cytology*; D050993:GATA6 Transcription Factor / Q000378:metabolism; D051379:Mice; D014212:Tretinoin / Q000378:metabolism
chemical_list: D050993:GATA6 Transcription Factor; D028341:Activins; D014212:Tretinoin
references: J Control Release. 1999 Aug 5;60(2-3):343-53
references: 10425339
references: Development. 1989 Apr;105(4):733-7
references: 2598811
references: Curr Stem Cell Res Ther. 2012 Jan;7(1):78-91
references: 22023624
references: Dev Biol. 1996 Jul 10;177(1):309-22
references: 8660897
references: Cell. 1980 Sep;21(2):347-55
references: 6250719
references: Nature. 1987 Dec 10-16;330(6148):576-8
references: 3683574
references: Vitam Horm. 2007;75:69-95
references: 17368312
references: Genesis. 2013 Apr;51(4):219-33
references: 23349011
references: Development. 2012 Aug;139(16):2866-77
references: 22791892
references: Dev Biol. 2010 Jan 1;337(1):63-73
references: 19850025
references: Angew Chem Int Ed Engl. 2003 Jul 14;42(27):3153-8
references: 12866105
references: Dev Biol. 2005 Oct 15;286(2):574-86
references: 16162334
references: BMC Dev Biol. 2007 Jul 03;7:80
references: 17605826
references: Methods Mol Med. 2005;105:23-46
references: 15492386
references: Stem Cells. 2002;20(2):139-45
references: 11897870
references: Stem Cells. 2012 Dec;30(12):2732-45
references: 23034951
references: Dev Biol. 2012 Jan 15;361(2):245-62
references: 22051107
references: Cell. 1995 Oct 20;83(2):279-87
references: 7585945
references: EMBO J. 2013 Apr 3;32(7):938-53
references: 23474895
references: Cell. 2006 Aug 25;126(4):663-76
references: 16904174
references: Nature. 2012 Jul 5;487(7405):57-63
references: 22722858
references: Proc Natl Acad Sci U S A. 1987 Nov;84(21):7413-7
references: 2823261
references: Pharm Res. 2007 Mar;24(3):438-49
references: 17252188
references: Cytotechnology. 2010 Jul;62(3):189-94
references: 20424915
references: Cell. 2005 Dec 2;123(5):917-29
references: 16325584
references: Nat Biotechnol. 2005 Dec;23(12):1542-50
references: 16311587
references: Nature. 1981 Jul 9;292(5819):154-6
references: 7242681
references: Dev Growth Differ. 2008 Sep;50(7):585-93
references: 19238729
references: Cell. 1987 Dec 24;51(6):987-1000
references: 3690668
references: Pharm Res. 2000 Feb;17(2):113-26
references: 10751024
references: Dev Cell. 2012 Apr 17;22(4):887-901
references: 22424930
references: Biotechniques. 1988 Jul-Aug;6(7):650-60
references: 3078721
references: Biotechniques. 1988 Sep;6(8):742-51
references: 3273636
references: PLoS One. 2010 Oct 20;5(10):e13446
references: 20975998
references: Development. 2010 Oct;137(20):3361-72
references: 20826533
references: Biol Open. 2012 Mar 15;1(3):208-19
references: 23213411
references: Cell. 2005 Aug 12;122(3):473-83
references: 16096065
references: Trends Genet. 1998 Jul;14(7):277-84
references: 9676530
references: Science. 1998 Dec 11;282(5396):2072-5
references: 9851926
references: Development. 2006 Jul;133(13):2497-505
references: 16728477
references: Development. 1999 Feb;126(4):723-32
references: 9895320
references: J Cell Sci. 2007 Nov 1;120(Pt 21):3859-69
references: 17940068
references: Proc Natl Acad Sci U S A. 1981 Dec;78(12):7634-8
references: 6950406
references: Dev Biol. 2012 Jan 1;361(1):90-102
references: 22027433
references: Dev Cell. 2008 Oct;15(4):509-20
references: 18854136
references: Cell Stem Cell. 2008 Nov 6;3(5):543-54
references: 18804426
references: Cell. 1999 Jan 22;96(2):195-209
references: 9988215
references: Curr Protoc Stem Cell Biol. 2008 Oct;Chapter 1:Unit 1E.4
references: 18972374
references: Gene Ther. 1995 Dec;2(10):710-22
references: 8750010
references: Genes Dev. 2010 Feb 1;24(3):312-26
references: 20123909
references: Development. 2005 Apr;132(7):1649-61
references: 15753215
references: Development. 2011 Sep;138(18):3885-95
references: 21862554
references: Stem Cells. 2011 Oct;29(10):1580-8
references: 22039605
references: Genes Dev. 2002 Apr 1;16(7):784-9
references: 11937486
references: Development. 2002 Aug;129(16):3913-24
references: 12135928
references: Nature. 1969 May 3;222(5192):480-1
references: 5768627
references: Biochim Biophys Acta. 1996 Jan 12;1278(1):41-50
references: 8611605
references: Development. 2013 Jan 15;140(2):267-79
references: 23193166
references: Cell Stem Cell. 2010 May 7;6(5):457-67
references: 20452320
references: Biotechniques. 1988 Oct;6(9):882-6
references: 3273199