title: Derivation conditions impact X-inactivation status in female human induced pluripotent stem cells.
abstract: Female human induced pluripotent stem cell (hiPSC) lines exhibit variability in X-inactivation status. The majority of hiPSC lines maintain one transcriptionally active X (Xa) and one inactive X (Xi) chromosome from donor cells. However, at low frequency, hiPSC lines with two Xas are produced, suggesting that epigenetic alterations of the Xi occur sporadically during reprogramming. We show here that X-inactivation status in female hiPSC lines depends on derivation conditions. hiPSC lines generated by the Kyoto method (retroviral or episomal reprogramming), which uses leukemia inhibitory factor (LIF)-expressing SNL feeders, frequently had two Xas. Early passage Xa/Xi hiPSC lines generated on non-SNL feeders were converted into Xa/Xa hiPSC lines after several passages on SNL feeders, and supplementation with recombinant LIF caused reactivation of some of X-linked genes. Thus, feeders are a significant factor affecting X-inactivation status. The efficient production of Xa/Xa hiPSC lines provides unprecedented opportunities to understand human X-reactivation and -inactivation.
authors: Gladstone Institute of Cardiovascular Disease, University of California, San Francisco, San Francisco, CA 94158, USA.
authors: Kiichiro
authors: K
authors: Tomoda
authors: Kazutoshi
authors: K
authors: Takahashi
authors: Karen
authors: K
authors: Leung
authors: Aki
authors: A
authors: Okada
authors: Megumi
authors: M
authors: Narita
authors: N Alice
authors: NA
authors: Yamada
authors: Kirsten E
authors: KE
authors: Eilertson
authors: Peter
authors: P
authors: Tsang
authors: Shiro
authors: S
authors: Baba
authors: Mark P
authors: MP
authors: White
authors: Salma
authors: S
authors: Sami
authors: Deepak
authors: D
authors: Srivastava
authors: Bruce R
authors: BR
authors: Conklin
authors: Barbara
authors: B
authors: Panning
authors: Shinya
authors: S
authors: Yamanaka
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D041321:Chromosomes, Human, X / Q000235:genetics; D061252:Feeder Cells / Q000166:cytology / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation; D050172:Genes, X-Linked; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D049951:X Chromosome Inactivation / Q000235:genetics*