title: Ascorbic acid prevents loss of Dlk1-Dio3 imprinting and facilitates generation of all-iPS cell mice from terminally differentiated B cells.
abstract: The generation of induced pluripotent stem cells (iPSCs) often results in aberrant epigenetic silencing of the imprinted Dlk1-Dio3 gene cluster, compromising the ability to generate entirely iPSC-derived adult mice ('all-iPSC mice'). Here, we show that reprogramming in the presence of ascorbic acid attenuates hypermethylation of Dlk1-Dio3 by enabling a chromatin configuration that interferes with binding of the de novo DNA methyltransferase Dnmt3a. This approach allowed us to generate all-iPSC mice from mature B cells, which have until now failed to support the development of exclusively iPSC-derived postnatal animals. Our data show that transcription factor-mediated reprogramming can endow a defined, terminally differentiated cell type with a developmental potential equivalent to that of embryonic stem cells. More generally, these findings indicate that culture conditions during cellular reprogramming can strongly influence the epigenetic and biological properties of the resultant iPSCs.
authors.affiliation: Massachusetts General Hospital Center for Regenerative Medicine, Harvard Stem Cell Institute, Boston, Massachusetts, USA.
authors.forename: Matthias
authors.initials: M
authors.lastname: Stadtfeld
authors.forename: Effie
authors.initials: E
authors.lastname: Apostolou
authors.forename: Francesco
authors.initials: F
authors.lastname: Ferrari
authors.forename: Jiho
authors.initials: J
authors.lastname: Choi
authors.forename: Ryan M
authors.initials: RM
authors.lastname: Walsh
authors.forename: Taiping
authors.initials: T
authors.lastname: Chen
authors.forename: Steen S K
authors.initials: SS
authors.lastname: Ooi
authors.forename: Sang Yong
authors.initials: SY
authors.lastname: Kim
authors.forename: Timothy H
authors.initials: TH
authors.lastname: Bestor
authors.forename: Toshi
authors.initials: T
authors.lastname: Shioda
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Park
authors.forename: Konrad
authors.initials: K
authors.lastname: Hochedlinger
mesh_terms: D000818:Animals; D001205:Ascorbic Acid / Q000494:pharmacology*; D001402:B-Lymphocytes / Q000166:cytology* / Q000378:metabolism*; D002135:Calcium-Binding Proteins; D002454:Cell Differentiation / Q000235:genetics; D065150:Cellular Reprogramming / Q000187:drug effects; D002843:Chromatin; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000378:metabolism; D019175:DNA Methylation / Q000187:drug effects; D000090205:DNA Methyltransferase 3A; D018392:Genomic Imprinting; D057026:Induced Pluripotent Stem Cells; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics*; D051379:Mice; D050814:Octamer Transcription Factor-3 / Q000235:genetics; D011401:Promoter Regions, Genetic; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D062085:RNA, Long Noncoding
chemical_list: D002135:Calcium-Binding Proteins; D002843:Chromatin; C512925:Dlk1 protein, mouse; C000715535:Dnmt3a protein, mouse; D036341:Intercellular Signaling Peptides and Proteins; C422882:MEG3 non-coding RNA, mouse; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D011506:Proteins; D062085:RNA, Long Noncoding; D004248:DNA (Cytosine-5-)-Methyltransferases; D000090205:DNA Methyltransferase 3A; D001205:Ascorbic Acid