TY - JOUR
T1 - Preventing erosion of X-chromosome inactivation in human embryonic stem cells
AU - Cloutier, Marissa
AU - Kumar, Surinder
AU - Buttigieg, Emily
AU - Keller, Laura
AU - Lee, Brandon
AU - Williams, Aaron
AU - Mojica-Perez, Sandra
AU - Erliandri, Indri
AU - Rocha, Andre Monteiro Da
AU - Cadigan, Kenneth
AU - Smith, Gary D.
AU - Kalantry, Sundeep
N1 - Funding Information:
We thank members of the Kalantry and Smith laboratories for discussions and critical review of the manuscript. This work was funded by NIH National Research Service Awards 5-T32-GM07544 (University of Michigan Predoctoral Genetics Training Program; to M.C.); T32-HD079342 (University of Michigan Predoctoral Career Training in the Reproductive Sciences Program; to M.C.); an NIH NIGMS R01 Award (R01GM124571) (to S. Kalantry); an NIH NICHD R01 Award (R01HD095463) (to S. Kalantry); a Reproductive Science Program Pilot Grant (to S. Ka. and G.D.S.); the University of Michigan Endowment for Basic Sciences (to S. Ka.); a University of Michigan Rackham Predoctoral Fellowship (to M.C.); MStem Cell Lab Funding (to G.D.S.); the University of Michigan President’s Office (to G.D.S.); Michigan Medicine (to G.D.S.); the A. Alfred Taubman Medical Research Institute (to G.D.S.); the University of Michigan Department of Obstetrics and Gynecology (to G.D.S.); and, the American Society for Reproductive Medicine (ASRM) Research Institute (to G.D.S.). We also thank Dr. Mark R. Hughes of Genesis Genetics for performing early passage hESC karyotyping by aCGH.
Funding Information:
We thank members of the Kalantry and Smith laboratories for discussions and critical review of the manuscript. This work was funded by NIH National Research Service Awards 5-T32-GM07544 (University of Michigan Predoctoral Genetics Training Program; to M.C.); T32-HD079342 (University of Michigan Predoctoral Career Training in the Reproductive Sciences Program; to M.C.); an NIH NIGMS R01 Award (R01GM124571) (to S. Kalantry); an NIH NICHD R01 Award (R01HD095463) (to S. Kalantry); a Reproductive Science Program Pilot Grant (to S. Ka. and G.D.S.); the University of Michigan Endowment for Basic Sciences (to S. Ka.); a University of Michigan Rackham Predoctoral Fellowship (to M.C.); MStem Cell Lab Funding (to G.D.S.); the University of Michigan President’s Office (to G.D.S.); Michigan Medicine (to G.D.S.); the A. Alfred Taubman Medical Research Institute (to G.D.S.); the University of Michigan Department of Obstetrics and Gynecology (to G.D.S.); and, the American Society for Reproductive Medicine (ASRM) Research Institute (to G.D.S.). We also thank Dr. Mark R. Hughes of Genesis Genetics for performing early passage hESC karyotyping by aCGH.
Publisher Copyright:
© 2022, The Author(s).
PY - 2022/12
Y1 - 2022/12
N2 - X-chromosome inactivation is a paradigm of epigenetic transcriptional regulation. Female human embryonic stem cells (hESCs) often undergo erosion of X-inactivation upon prolonged culture. Here, we investigate the sources of X-inactivation instability by deriving new primed pluripotent hESC lines. We find that culture media composition dramatically influenced the expression of XIST lncRNA, a key regulator of X-inactivation. hESCs cultured in a defined xenofree medium stably maintained XIST RNA expression and coating, whereas hESCs cultured in the widely used mTeSR1 medium lost XIST RNA expression. We pinpointed lithium chloride in mTeSR1 as a cause of XIST RNA loss. The addition of lithium chloride or inhibitors of GSK-3 proteins that are targeted by lithium to the defined hESC culture medium impeded XIST RNA expression. GSK-3 inhibition in differentiating female mouse embryonic stem cells and epiblast stem cells also resulted in a loss of XIST RNA expression. Together, these data may reconcile observed variations in X-inactivation in hESCs and inform the faithful culture of pluripotent stem cells.
AB - X-chromosome inactivation is a paradigm of epigenetic transcriptional regulation. Female human embryonic stem cells (hESCs) often undergo erosion of X-inactivation upon prolonged culture. Here, we investigate the sources of X-inactivation instability by deriving new primed pluripotent hESC lines. We find that culture media composition dramatically influenced the expression of XIST lncRNA, a key regulator of X-inactivation. hESCs cultured in a defined xenofree medium stably maintained XIST RNA expression and coating, whereas hESCs cultured in the widely used mTeSR1 medium lost XIST RNA expression. We pinpointed lithium chloride in mTeSR1 as a cause of XIST RNA loss. The addition of lithium chloride or inhibitors of GSK-3 proteins that are targeted by lithium to the defined hESC culture medium impeded XIST RNA expression. GSK-3 inhibition in differentiating female mouse embryonic stem cells and epiblast stem cells also resulted in a loss of XIST RNA expression. Together, these data may reconcile observed variations in X-inactivation in hESCs and inform the faithful culture of pluripotent stem cells.
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UR - http://www.scopus.com/inward/citedby.url?scp=85129757077&partnerID=8YFLogxK
U2 - 10.1038/s41467-022-30259-x
DO - 10.1038/s41467-022-30259-x
M3 - Article
C2 - 35523820
AN - SCOPUS:85129757077
SN - 2041-1723
VL - 13
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 2516
ER -