title: Partial reversal of the methylation pattern of the X-linked gene HUMARA during hematopoietic differentiation of human embryonic stem cells.
abstract: Human embryonic stem cells (hESCs) can be induced to differentiate towards hematopoiesis with high efficiency. In this work, we analyzed the methylation status of the X-linked HUMARA (human androgen receptor) gene in hematopoietic cells derived from hESC line H9 before and after induction of hematopoietic differentiation. All passages of H9 and H9-derived hematopoietic cells displayed homogenous methylation pattern with disappearance of the same allele upon HpaII digestion. This pattern persisted in the great majority of different hematopoietic progenitors derived from H9, except in 11 of 86 individually plucked colonies in which an equal digestion of the HUMARA alleles has been found, suggesting that a methylation change occurring at this locus during differentiation. Interestingly, quantification of X inactive-specific transcript (XIST) RNA in undifferentiated H9 cell line and day 14 embryoid bodies (EB) by RT-PCR did not show any evidence of XIST expression either before or after differentiation. Thus, during self-renewal conditions and after induction of commitment towards the formation of EB, the methylation pattern of the HUMARA locus appears locked with the same unmethylated allele. However, hematopoietic differentiation seems to be permissive to the reversal of methylation status of HUMARA in some terminally differentiated progenitors. These data suggest that monitoring methylation of HUMARA gene during induced differentiation could be of use for studying hESC-derived hematopoiesis.
authors.affiliation: INSERM U935, Université Paris Sud 11, AP-HP, Villejuif 94802, France.
authors.forename: Maria-Teresa
authors.initials: MT
authors.lastname: Mitjavila-Garcia
authors.forename: Marie Laure
authors.initials: ML
authors.lastname: Bonnet
authors.forename: Frank
authors.initials: F
authors.lastname: Yates
authors.forename: Rima
authors.initials: R
authors.lastname: Haddad
authors.forename: Noufissa
authors.initials: N
authors.lastname: Oudrhiri
authors.forename: Olivier
authors.initials: O
authors.lastname: Féraud
authors.forename: Aurelie
authors.initials: A
authors.lastname: Magniez
authors.forename: Melanie
authors.initials: M
authors.lastname: Makhlouf
authors.forename: Celine
authors.initials: C
authors.lastname: Vallot
authors.forename: Claire
authors.initials: C
authors.lastname: Rougeulle
authors.forename: Annelise
authors.initials: A
authors.lastname: Bennaceur-Griscelli
authors.forename: Ali G
authors.initials: AG
authors.lastname: Turhan
mesh_terms: D002460:Cell Line; D019175:DNA Methylation; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D050172:Genes, X-Linked; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D011944:Receptors, Androgen / Q000235:genetics* / Q000378:metabolism
chemical_list: C410798:AR protein, human; D011944:Receptors, Androgen