Derivation of pre-X inactivation human embryonic stem cells under physiological oxygen concentrations.
The presence of two active X chromosomes (XaXa) is a hallmark of the ground state of pluripotency specific to murine embryonic stem cells (ESCs). Human ESCs (hESCs) invariably exhibit signs of X chromosome inactivation (XCI) and are considered developmentally more advanced than their murine counterparts. We describe the establishment of XaXa hESCs derived under physiological oxygen concentrations. Using these cell lines, we demonstrate that (1) differentiation of hESCs induces random XCI in a manner similar to murine ESCs, (2) chronic exposure to atmospheric oxygen is sufficient to induce irreversible XCI with minor changes of the transcriptome, (3) the Xa exhibits heavy methylation of the XIST promoter region, and (4) XCI is associated with demethylation and transcriptional activation of XIST along with H3K27-me3 deposition across the Xi. These findings indicate that the human blastocyst contains pre-X-inactivation cells and that this state is preserved in vitro through culture under physiological oxygen.
Lengner
Christopher J
CJ
Whitehead Institute for Biomedical Sciences, 9 Cambridge Center, Cambridge, MA 02142, USA.
Gimelbrant
Alexander A
AA
Erwin
Jennifer A
JA
Cheng
Albert Wu
AW
Guenther
Matthew G
MG
Welstead
G Grant
GG
Alagappan
Raaji
R
Frampton
Garrett M
GM
Xu
Ping
P
Muffat
Julien
J
Santagata
Sandro
S
Powers
Doug
D
Barrett
C Brent
CB
Young
Richard A
RA
Lee
Jeannie T
JT
Jaenisch
Rudolf
R
Mitalipova
Maisam
M
D000818:Animals; D002454:Cell Differentiation; D041321:Chromosomes, Human, X / Q000378:metabolism*; D053595:Embryonic Stem Cells / Q000378:metabolism*; D005260:Female; D006657:Histones / Q000378:metabolism; D006801:Humans; D007621:Karyotyping; D008297:Male; D051379:Mice; D018384:Oxidative Stress; D010100:Oxygen / Q000378:metabolism*; D039904:Pluripotent Stem Cells / Q000378:metabolism; D049951:X Chromosome Inactivation
D006657:Histones; D010100:Oxygen