title: Stable X chromosome reactivation in female human induced pluripotent stem cells.
abstract: In placental mammals, balanced expression of X-linked genes is accomplished by X chromosome inactivation (XCI) in female cells. In humans, random XCI is initiated early during embryonic development. To investigate whether reprogramming of female human fibroblasts into induced pluripotent stem cells (iPSCs) leads to reactivation of the inactive X chromosome (Xi), we have generated iPSC lines from fibroblasts heterozygous for large X-chromosomal deletions. These fibroblasts show completely skewed XCI of the mutated X chromosome, enabling monitoring of X chromosome reactivation (XCR) and XCI using allele-specific single-cell expression analysis. This approach revealed that XCR is robust under standard culture conditions, but does not prevent reinitiation of XCI, resulting in a mixed population of cells with either two active X chromosomes (Xas) or one Xa and one Xi. This mixed population of XaXa and XaXi cells is stabilized in naive human stem cell medium, allowing expansion of clones with two Xas.
authors: Department of Developmental Biology, Erasmus MC, University Medical Center, 3015 CE Rotterdam, the Netherlands.
authors: Tahsin Stefan
authors: TS
authors: Barakat
authors: Department of Developmental Biology, Erasmus MC, University Medical Center, 3015 CE Rotterdam, the Netherlands; Erasmus Stem Cell and Regenerative Medicine Institute, Erasmus MC, University Medical Center, 3015 CE Rotterdam, the Netherlands.
authors: Mehrnaz
authors: M
authors: Ghazvini
authors: Department of Developmental Biology, Erasmus MC, University Medical Center, 3015 CE Rotterdam, the Netherlands; Department of Obstetrics and Gynecology, Erasmus MC, University Medical Center, 3015 CE Rotterdam, the Netherlands.
authors: Bas
authors: B
authors: de Hoon
authors: Department of Developmental Biology, Erasmus MC, University Medical Center, 3015 CE Rotterdam, the Netherlands; Erasmus Stem Cell and Regenerative Medicine Institute, Erasmus MC, University Medical Center, 3015 CE Rotterdam, the Netherlands.
authors: Tracy
authors: T
authors: Li
authors: Department of Clinical Genetics, Erasmus MC, University Medical Center, 3015 CE Rotterdam, the Netherlands.
authors: Bert
authors: B
authors: Eussen
authors: Department of Clinical Genetics, Erasmus MC, University Medical Center, 3015 CE Rotterdam, the Netherlands.
authors: Hannie
authors: H
authors: Douben
authors: Erasmus Stem Cell and Regenerative Medicine Institute, Erasmus MC, University Medical Center, 3015 CE Rotterdam, the Netherlands.
authors: Reinier
authors: R
authors: van der Linden
authors: Department of Developmental Biology, Erasmus MC, University Medical Center, 3015 CE Rotterdam, the Netherlands; Erasmus Stem Cell and Regenerative Medicine Institute, Erasmus MC, University Medical Center, 3015 CE Rotterdam, the Netherlands.
authors: Nathalie
authors: N
authors: van der Stap
authors: Department of Clinical Genetics, Erasmus MC, University Medical Center, 3015 CE Rotterdam, the Netherlands.
authors: Marjan
authors: M
authors: Boter
authors: Department of Obstetrics and Gynecology, Erasmus MC, University Medical Center, 3015 CE Rotterdam, the Netherlands.
authors: Joop S
authors: JS
authors: Laven
authors: Department of Clinical Genetics, Erasmus MC, University Medical Center, 3015 CE Rotterdam, the Netherlands.
authors: Robert-Jan
authors: RJ
authors: Galjaard
authors: Department of Developmental Biology, Erasmus MC, University Medical Center, 3015 CE Rotterdam, the Netherlands.
authors: J Anton
authors: JA
authors: Grootegoed
authors: Department of Clinical Genetics, Erasmus MC, University Medical Center, 3015 CE Rotterdam, the Netherlands.
authors: Annelies
authors: A
authors: de Klein
authors: Department of Developmental Biology, Erasmus MC, University Medical Center, 3015 CE Rotterdam, the Netherlands. Electronic address: j.gribnau@erasmusmc.nl.
authors: Joost
authors: J
authors: Gribnau
mesh_terms: D002460:Cell Line; D002478:Cells, Cultured; D002874:Chromosome Mapping; D041321:Chromosomes, Human, X; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D015870:Gene Expression; D023061:Gene Order; D050172:Genes, X-Linked; D056426:Genetic Loci; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D059785:Karyotype; D015533:Transcriptional Activation; D019076:Transgenes; D049951:X Chromosome Inactivation
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