Derivation Conditions Impact X-Inactivation Status in Female Human Induced Pluripotent Stem Cells

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, sugge...

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Bibliographic Details
Published in:Cell stem cell 2012-07, Vol.11 (1), p.91-99
Main Authors: Tomoda, Kiichiro, Takahashi, Kazutoshi, Leung, Karen, Okada, Aki, Narita, Megumi, Yamada, N. Alice, Eilertson, Kirsten E., Tsang, Peter, Baba, Shiro, White, Mark P., Sami, Salma, Srivastava, Deepak, Conklin, Bruce R., Panning, Barbara, Yamanaka, Shinya
Format: Article
Language:English
Subjects:
Biological and medical sciences
Cell Culture Techniques - methods
Cell Differentiation - genetics
Cell differentiation, maturation, development, hematopoiesis
Cell Line
Cell physiology
Chromosomes, Human, X - genetics
Feeder Cells - cytology
Feeder Cells - metabolism
Female
Fundamental and applied biological sciences. Psychology
Gene Expression Regulation
Genes, X-Linked
Humans
Induced Pluripotent Stem Cells - cytology
Induced Pluripotent Stem Cells - metabolism
Molecular and cellular biology
Sequence Analysis, DNA
X Chromosome Inactivation - genetics
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Summary: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. [Display omitted] ► X-Inactivation status in female hiPSC lines depends on derivation conditions ► hiPSC lines generated by the Kyoto method on SNL feeders frequently had two Xas ► Leukemia inhibitory factor (LIF) is a key factor in X-reactivation ► Xa/Xa hiPSC lines are valuable tools for understanding human X-inactivation Growth conditions, particularly the presence of SNL feeder cells and LIF, strongly influence the degree of X-reactivation seen in female hiPSCs. This discovery helps clarify previous confusion about the X-inactivation status of female hiPSCs and provides a tool for analysis of human X-inactivation mechanisms.
ISSN:1934-5909
1875-9777
DOI:10.1016/j.stem.2012.05.019