Isolation of Oct4-Expressing Extraembryonic Endoderm Precursor Cell Lines

被引:40
作者
Debeb, Bisrat G.
Galat, Vasiliy
Epple-Farmer, Jessica
Iannaccone, Steve
Woodward, Wendy A.
Bader, Michael
Iannaccone, Philip
Binas, Bert
机构
[1] Department of Pathobiology, College of Veterinary Medicine and Biomedical Sciences, Texas AandM University, College Station, TX
[2] Developmental Biology Program, Children's Memorial Research Center, Northwestern University, Chicago, IL
[3] Division of Radiation Oncology, University of Texas M.D. Anderson Cancer Center, Houston, TX
[4] Max Delbruck Center for Molecular Medicine, Berlin
[5] Division of Molecular and Life Sciences, Hanyang University, Ansan-si, Gyeonggi-do
来源
PLOS ONE | 2009年 / 4卷 / 09期
关键词
D O I
10.1371/journal.pone.0007216
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Background: The extraembryonic endoderm (ExEn) defines the yolk sac, a set of membranes that provide essential support for mammalian embryos. Recent findings suggest that the committed ExEn precursor is present already in the embryonic Inner Cell Mass (ICM) as a group of cells that intermingles with the closely related epiblast precursor. All ICM cells contain Oct4, a key transcription factor that is first expressed at the morula stage. In vitro, the epiblast precursor is most closely represented by the well-characterized embryonic stem (ES) cell lines that maintain the expression of Oct4, but analogous ExEn precursor cell lines are not known and it is unclear if they would express Oct4. Methodology/Principal Findings: Here we report the isolation and characterization of permanently proliferating Oct4-expressing rat cell lines ("XEN-P cell lines''), which closely resemble the ExEn precursor. We isolated the XEN-P cell lines from blastocysts and characterized them by plating and gene expression assays as well as by injection into embryos. Like ES cells, the XEN-P cells express Oct4 and SSEA1 at high levels and their growth is stimulated by leukemia inhibitory factor, but instead of the epiblast determinant Nanog, they express the ExEn determinants Gata6 and Gata4. Further, they lack markers characteristic of the more differentiated primitive/visceral and parietal ExEn stages, but exclusively differentiate into these stages in vitro and contribute to them in vivo. Conclusions/Significance: Our findings (i) suggest strongly that the ExEn precursor is a self-renewable entity, (ii) indicate that active Oct4 gene expression (transcription plus translation) is part of its molecular identity, and (iii) provide an in vitro model of early ExEn differentiation.
引用
收藏
页数:13
相关论文
共 58 条
[1]  
[Anonymous], 2012, Molecular Cloning: A Laboratory Manual
[2]   Embryonic origins of mammalian hematopoiesis [J].
Baron, MH .
EXPERIMENTAL HEMATOLOGY, 2003, 31 (12) :1160-1169
[3]   Parameters Influencing Derivation of Embryonic Stem Cells From Murine Embryos [J].
Batlle-Morera, Laura ;
Smith, Austin ;
Nichols, Jennifer .
GENESIS, 2008, 46 (12) :758-767
[4]   Derivation of pluripotent epiblast stem cells from mammalian embryos [J].
Brons, I. Gabrielle M. ;
Smithers, Lucy E. ;
Trotter, Matthew W. B. ;
Rugg-Gunn, Peter ;
Sun, Bowen ;
de Sousa Lopes, Susana M. Chuva ;
Howlett, Sarah K. ;
Clarkson, Amanda ;
Ahrlund-Richter, Lars ;
Pedersen, Roger A. ;
Vallier, Ludovic .
NATURE, 2007, 448 (7150) :191-U7
[5]   Rapid loss of Oct-4 and pluripotency in cultured rodent blastocysts and derivative cell lines [J].
Buehr, M ;
Nichols, J ;
Stenhouse, F ;
Mountford, P ;
Greenhalgh, CJ ;
Kantachuvesiri, S ;
Brooker, G ;
Mullins, J ;
Smith, AG .
BIOLOGY OF REPRODUCTION, 2003, 68 (01) :222-229
[6]   Capture of Authentic Embryonic Stem Cells from Rat Blastocysts [J].
Buehr, Mia ;
Meek, Stephen ;
Blair, Kate ;
Yang, Jian ;
Ure, Janice ;
Silva, Jose ;
McLay, Renee ;
Hall, John ;
Ying, Qi-Long ;
Smith, Austin .
CELL, 2008, 135 (07) :1287-1298
[7]   Nutrition of the human fetus during the first trimester - A review [J].
Burton, GJ ;
Hempstock, J ;
Jauniaux, E .
PLACENTA, 2001, 22 :S70-S76
[8]   Self-renewal of teratocarcinoma and embryonic stem cells [J].
Chambers, I ;
Smith, A .
ONCOGENE, 2004, 23 (43) :7150-7160
[9]  
CHAZAUD C, 2006, DEV CELL, V615, P24
[10]   Expression of the T-box gene Eomesodermin during early mouse development [J].
Ciruna, BG ;
Rossant, J .
MECHANISMS OF DEVELOPMENT, 1999, 81 (1-2) :199-203