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 language | English |
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Pages (from-to) | 276-285 |
Number of pages | 10 |
Journal | Stem cell reviews and reports |
Volume | 15 |
Issue number | 2 |
Early online date | 13 Nov 2018 |
DOIs | |
Publication status | Published - 15 Apr 2019 |
Keywords
- Episomal reprogramming
- Isogenic control
- Rett syndrome
- X-chromosome
- iPSCs