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 - 2019/4/15
Y1 - 2019/4/15
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.
KW - Episomal reprogramming
KW - Isogenic control
KW - Rett syndrome
KW - X-chromosome
KW - iPSCs
UR - http://www.scopus.com/inward/record.url?scp=85056375035&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85056375035&partnerID=8YFLogxK
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UR - https://www.ncbi.nlm.nih.gov/pubmed/30421281
U2 - https://doi.org/10.1007/s12015-018-9851-8
DO - https://doi.org/10.1007/s12015-018-9851-8
M3 - Article
C2 - 30421281
SN - 1550-8943
VL - 15
SP - 276
EP - 285
JO - STEM CELL REVIEWS AND REPORTS
JF - STEM CELL REVIEWS AND REPORTS
IS - 2
ER -