title: Valproic acid confers functional pluripotency to human amniotic fluid stem cells in a transgene-free approach.
abstract: Induced pluripotent stem cells (iPSCs) with potential for therapeutic applications can be derived from somatic cells via ectopic expression of a set of limited and defined transcription factors. However, due to risks of random integration of the reprogramming transgenes into the host genome, the low efficiency of the process, and the potential risk of virally induced tumorigenicity, alternative methods have been developed to generate pluripotent cells using nonintegrating systems, albeit with limited success. Here, we show that c-KIT+ human first-trimester amniotic fluid stem cells (AFSCs) can be fully reprogrammed to pluripotency without ectopic factors, by culture on Matrigel in human embryonic stem cell (hESC) medium supplemented with the histone deacetylase inhibitor (HDACi) valproic acid (VPA). The cells share 82% transcriptome identity with hESCs and are capable of forming embryoid bodies (EBs) in vitro and teratomas in vivo. After long-term expansion, they maintain genetic stability, protein level expression of key pluripotency factors, high cell-division kinetics, telomerase activity, repression of X-inactivation, and capacity to differentiate into lineages of the three germ layers, such as definitive endoderm, hepatocytes, bone, fat, cartilage, neurons, and oligodendrocytes. We conclude that AFSC can be utilized for cell banking of patient-specific pluripotent cells for potential applications in allogeneic cellular replacement therapies, pharmaceutical screening, and disease modeling.
authors: Imperial College London, London, UK.
authors: Dafni
authors: D
authors: Moschidou
authors: Sayandip
authors: S
authors: Mukherjee
authors: Michael P
authors: MP
authors: Blundell
authors: Katharina
authors: K
authors: Drews
authors: Gemma N
authors: GN
authors: Jones
authors: Hassan
authors: H
authors: Abdulrazzak
authors: Beata
authors: B
authors: Nowakowska
authors: Anju
authors: A
authors: Phoolchund
authors: Kenneth
authors: K
authors: Lay
authors: T Selvee
authors: TS
authors: Ramasamy
authors: Mara
authors: M
authors: Cananzi
authors: Daniel
authors: D
authors: Nettersheim
authors: Mark
authors: M
authors: Sullivan
authors: Jennifer
authors: J
authors: Frost
authors: Gudrun
authors: G
authors: Moore
authors: Joris R
authors: JR
authors: Vermeesch
authors: Nicholas M
authors: NM
authors: Fisk
authors: Adrian J
authors: AJ
authors: Thrasher
authors: Anthony
authors: A
authors: Atala
authors: James
authors: J
authors: Adjaye
authors: Hubert
authors: H
authors: Schorle
authors: Paolo
authors: P
authors: De Coppi
authors: Pascale V
authors: PV
authors: Guillot
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology / Q000187:drug effects*; D002454:Cell Differentiation; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D005260:Female; D015894:Genome, Human; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects* / Q000378:metabolism; D007621:Karyotyping; D007700:Kinetics; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000235:genetics / Q000378:metabolism; D008297:Male; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D059467:Transcriptome; D019076:Transgenes; D014635:Valproic Acid / Q000494:pharmacology*; D049951:X Chromosome Inactivation / Q000187:drug effects
chemical_list: D004268:DNA-Binding Proteins; D056572:Histone Deacetylase Inhibitors; D018398:Homeodomain Proteins; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; C116115:MYCBP protein, human; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; D014157:Transcription Factors; D014635:Valproic Acid