TY - JOUR
T1 - Totipotent Embryonic Stem Cells Arise in Ground-State Culture Conditions
AU - Morgani, Sophie M.
AU - Canham, Maurice A.
AU - Nichols, Jennifer
AU - Sharov, Alexei A.
AU - Migueles, Rosa Portero
AU - Ko, Minoru S.H.
AU - Brickman, Joshua M.
N1 - Funding Information:
We thank Lynsey Robertson, Sally Inverarity, Javier Martin Gonzalez, Kasper Bonderup, Ron Wilkie, Simon Monard, Gelo de la Cruz, and Valerie Wilson for technical assistance; Austin Smith, Heiko Lickert, and Ian Chambers for reagents; and Sophie Astrof, Sally Lowell, and the entire Brickman lab for critical discussion of this manuscript. This work was supported by grants from the Medical Research Council (MRC) (G0701428) and the Novo Nordisk Foundation (to J.M.B.), the Intramural Research Program of the National Institutes of Health, and the National Institute on Aging (Z01AG AG000656 and Z01AG000662 to M.S.H.K.). S.M.M. is supported by an MRC studentship, and J.M.B. is an MRC senior nonclinical fellow. The funders had no role in study design, data collection, and analysis, decision to publish, or preparation of the manuscript. S.M.M. conceived of and executed experiments and wrote the paper with J.M.B.; M.A.C. generated the HV mice and ESCs; J.N. designed and executed the experiments and helped with embryo culture; A.A.S. analyzed RNA-seq data sets; R.P.M. executed experiments; M.S.H.K. supervised data analysis; and J.M.B. designed and supervised experiments and wrote the paper with S.M.M.
PY - 2013/3/27
Y1 - 2013/3/27
N2 - Embryonic stem cells (ESCs) are derived from mammalian embryos during the transition from totipotency, when individual blastomeres can make all lineages, to pluripotency, when they are competent to make only embryonic lineages. ESCs maintained with inhibitors of MEK and GSK3 (2i) are thought to represent an embryonically restricted ground state. However, we observed heterogeneous expression of the extraembryonic endoderm marker Hex in 2i-cultured embryos, suggesting that 2i blocked development prior to epiblast commitment. Similarly, 2i ESC cultures were heterogeneous and contained a Hex-positive fraction primed to differentiate into trophoblast and extraembryonic endoderm. Single Hex-positive ESCs coexpressed epiblast and extraembryonic genes and contributed to all lineages in chimeras. The cytokine LIF, necessary for ESC self-renewal, supported the expansion of this population but did not directly support Nanog-positive epiblast-like ESCs. Thus, 2i and LIF support a totipotent state comparable to early embryonic cells that coexpress embryonic and extraembryonic determinants.
AB - Embryonic stem cells (ESCs) are derived from mammalian embryos during the transition from totipotency, when individual blastomeres can make all lineages, to pluripotency, when they are competent to make only embryonic lineages. ESCs maintained with inhibitors of MEK and GSK3 (2i) are thought to represent an embryonically restricted ground state. However, we observed heterogeneous expression of the extraembryonic endoderm marker Hex in 2i-cultured embryos, suggesting that 2i blocked development prior to epiblast commitment. Similarly, 2i ESC cultures were heterogeneous and contained a Hex-positive fraction primed to differentiate into trophoblast and extraembryonic endoderm. Single Hex-positive ESCs coexpressed epiblast and extraembryonic genes and contributed to all lineages in chimeras. The cytokine LIF, necessary for ESC self-renewal, supported the expansion of this population but did not directly support Nanog-positive epiblast-like ESCs. Thus, 2i and LIF support a totipotent state comparable to early embryonic cells that coexpress embryonic and extraembryonic determinants.
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U2 - 10.1016/j.celrep.2013.04.034
DO - 10.1016/j.celrep.2013.04.034
M3 - Article
C2 - 23746443
AN - SCOPUS:84879796179
VL - 3
SP - 1945
EP - 1957
JO - Cell Reports
JF - Cell Reports
SN - 2211-1247
IS - 6
ER -