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 |
|---|---|
| Journal | Stem cell reviews and reports |
| DOIs | |
| Publication status | E-pub ahead of print - 13 Nov 2018 |
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Generation of Isogenic Controls for In Vitro Disease Modelling of X-Chromosomal Disorders. / Hinz, Lisa; Hoekstra, Stephanie D; Watanabe, Kyoko; Posthuma, Danielle; Heine, Vivi M.
In: Stem cell reviews and reports, 13.11.2018.Research output: Contribution to Journal › Article › Academic › peer-review
TY - JOUR
T1 - Generation of Isogenic Controls for In Vitro Disease Modelling of X-Chromosomal Disorders
AU - Hinz, Lisa
AU - Hoekstra, Stephanie D
AU - Watanabe, Kyoko
AU - Posthuma, Danielle
AU - Heine, Vivi M
PY - 2018/11/13
Y1 - 2018/11/13
N2 - 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.
AB - 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.
U2 - 10.1007/s12015-018-9851-8
DO - 10.1007/s12015-018-9851-8
M3 - Article
JO - Stem cell reviews and reports
JF - Stem cell reviews and reports
SN - 1550-8943
ER -