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

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

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
Volume15
Issue number2
Early online date13 Nov 2018
DOIs
Publication statusPublished - 15 Apr 2019

Keywords

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

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