Generation of Isogenic Controls for In Vitro Disease Modelling of X-Chromosomal Disorders

Lisa Hinz, Stephanie D Hoekstra, Kyoko Watanabe, Danielle Posthuma, Vivi M Heine

Research output: Contribution to JournalArticleAcademicpeer-review


Generation of proper controls is crucial in induced pluripotent stem cell (iPSC) studies. X-chromosomal disorders offer the potential to develop isogenic controls due to random X-chromosomal inactivation (XCI). However, the generation of such lines is currently hampered by skewed X-inactivation in fibroblast lines and X-chromosomal reactivation (XCR) after reprogramming. Here we describe a method to generate a pure iPSC population with respect to the specific inactivated X-chromosome (Xi). We used fibroblasts from Rett patients, who all have a causal mutation in the X-linked MeCP2 gene. Pre-sorting these fibroblasts followed by episomal reprogramming, allowed us to overcome skewness in fibroblast lines and to retain the X-chromosomal state, which was unpredictable with lentiviral reprogramming. This means that fibroblast pre-sorting followed by episomal reprogramming can be used to reliably generate iPSC lines with specified X-chromosomal phenotype such as Rett syndrome.

Original languageEnglish
Pages (from-to)276-285
Number of pages10
JournalStem cell reviews and reports
Issue number2
Early online date13 Nov 2018
Publication statusPublished - 15 Apr 2019


Acknowledgements Specimens were provided by the Cell lines and DNA bank of Rett Syndrome, X-linked mental retardation and other genetic disease, member of the Telethon Network of Genetic Biobanks (project no. GTB12001), funded by Telethon Italy, and of the EuroBioBank network. Funding This work was funded by the EC under FP7-PEOPLE-2013 (607508) and The Netherlands Organization for Scientific Research (NWO VICI 453–14-005). VMH is supported by ZonMw (VIDI 917– 12-343).

FundersFunder number
EuroBioBank network
Netherlands Organization for Scientific Research
Telethon Italy
Seventh Framework Programme607508
European CommissionFP7-PEOPLE-2013
ZonMwVIDI 917– 12-343
Nederlandse Organisatie voor Wetenschappelijk OnderzoekVICI 453–14-005


    • Episomal reprogramming
    • Isogenic control
    • Rett syndrome
    • X-chromosome
    • iPSCs


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