Imprinting fidelity in mouse iPSCs depends on sex of donor cell and medium formulation

Autor: Arez, Maria, Eckersley-Maslin, Melanie, Klobuar, Tajda, von Gilsa Lopes, João, Krueger, Felix, Mupo, Annalisa, Raposo, Ana Cláudia, Oxley, David, Mancino, Samantha, Gendrel, Anne-Valerie, Jesus, Bruno Bernardes De, da Rocha, Simão T.
Přispěvatelé: NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM), Repositório da Universidade de Lisboa
Rok vydání: 2022
Předmět:
Zdroj: Nature Communications. 13
ISSN: 2041-1723
DOI: 10.1038/s41467-022-33013-5
Popis: © The Author(s) 2022 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
Reprogramming of somatic cells into induced Pluripotent Stem Cells (iPSCs) is a major leap towards personalised approaches to disease modelling and cell-replacement therapies. However, we still lack the ability to fully control the epigenetic status of iPSCs, which is a major hurdle for their downstream applications. Epigenetic fidelity can be tracked by genomic imprinting, a phenomenon dependent on DNA methylation, which is frequently perturbed in iPSCs by yet unknown reasons. To try to understand the causes underlying these defects, we conducted a thorough imprinting analysis using IMPLICON, a high-throughput method measuring DNA methylation levels, in multiple female and male murine iPSC lines generated under different experimental conditions. Our results show that imprinting defects are remarkably common in iPSCs, but their nature depends on the sex of donor cells and their response to culture conditions. Imprints in female iPSCs resist the initial genome-wide DNA demethylation wave during reprogramming, but ultimately cells accumulate hypomethylation defects irrespective of culture medium formulations. In contrast, imprinting defects on male iPSCs depends on the experimental conditions and arise during reprogramming, being mitigated by the addition of vitamin C (VitC). Our findings are fundamental to further optimise reprogramming strategies and generate iPSCs with a stable epigenome.
Work in S.T.d.R.’s team was supported by Fundação para a Ciência e Tecnologia (FCT) Ministério da Ciência, Tecnologia e Ensino Superior (MCTES), Portugal [IC&DT projects PTDC/BEX-BCM/2612/2014 and PTDC/BIA-MOL/29320/2017 as well as projects UIDB/04565/2020 and UIDP/04565/2020 of the Research Unit Institute from Bioengineering and Biosciences – iBB and LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy – i4HB]; S.T.d.R. and A.-V.G. are supported by assistant research contracts from FCT/MCTES (CEECIND/01234/2017 and CEECIND/02085/2018, respectively); M.A and A.C.R. are supported, respectively, by SFRH/BD/151251/2021 and SFRH/BD/137099/2018 PhD fellowships from FCT/MCTES. J.V.G.L is supported by COVID/BD/152624/2022 from FCT/MCTES. MAE-M was supported by a BBSRC Discovery Fellowship (BB/T009713/1) and is now supported by a Snow Medical Fellowship. F.K. is supported by the Babraham Institute Strategic Core Funding and A.M. by BBSRC BBS/E/B/000C0421. B.B.J. work was funded by Fundação para a Ciência e Tecnologia (FCT), and FEDER, LISBOA-01-0145-FEDER-028534, project co-funded by FEDER, through POR Lisboa 2020—Programa Operacional Regional de Lisboa. T.K. is supported by Janko Jamnik Doctoral Scholarship from National Institute of Chemistry.
Databáze: OpenAIRE
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