title: Trisomy 21 mid-trimester amniotic fluid induced pluripotent stem cells maintain genetic signatures during reprogramming: implications for disease modeling and cryobanking.
abstract: Trisomy 21 is the most common chromosomal abnormality and is associated primarily with cardiovascular, hematological, and neurological complications. A robust patient-derived cellular model is necessary to investigate the pathophysiology of the syndrome because current animal models are limited and access to tissues from affected individuals is ethically challenging. We aimed to derive induced pluripotent stem cells (iPSCs) from trisomy 21 human mid-trimester amniotic fluid stem cells (AFSCs) and describe their hematopoietic and neurological characteristics. Human AFSCs collected from women undergoing prenatal diagnosis were selected for c-KIT(+) and transduced with a Cre-lox-inducible polycistronic lentiviral vector encoding SOX2, OCT4, KLF-4, and c-MYC (50,000 cells at a multiplicity of infection (MOI) 1-5 for 72 h). The embryonic stem cell (ESC)-like properties of the AFSC-derived iPSCs were established in vitro by embryoid body formation and in vivo by teratoma formation in RAG2(-/-), γ-chain(-/-), C2(-/-) immunodeficient mice. Reprogrammed cells retained their cytogenetic signatures and differentiated into specialized hematopoietic and neural precursors detected by morphological assessment, immunostaining, and RT-PCR. Additionally, the iPSCs expressed all pluripotency markers upon multiple rounds of freeze-thawing. These findings are important in establishing a patient-specific cellular platform of trisomy 21 to study the pathophysiology of the aneuploidy and for future drug discovery.
authors.affiliation: 1 Surgery Unit, Institute of Child Health, University College London , London, WC1N 1EH, United Kingdom .
authors.forename: Caterina
authors.initials: C
authors.lastname: Pipino
authors.forename: Sayandip
authors.initials: S
authors.lastname: Mukherjee
authors.forename: Anna L
authors.initials: AL
authors.lastname: David
authors.forename: Michael P
authors.initials: MP
authors.lastname: Blundell
authors.forename: Steven W
authors.initials: SW
authors.lastname: Shaw
authors.forename: Peggy
authors.initials: P
authors.lastname: Sung
authors.forename: Panicos
authors.initials: P
authors.lastname: Shangaris
authors.forename: Jonathan J
authors.initials: JJ
authors.lastname: Waters
authors.forename: Drew
authors.initials: D
authors.lastname: Ellershaw
authors.forename: Marina
authors.initials: M
authors.lastname: Cavazzana
authors.forename: Gustavo
authors.initials: G
authors.lastname: Mostoslavsky
authors.forename: Assunta
authors.initials: A
authors.lastname: Pandolfi
authors.forename: Agostino
authors.initials: A
authors.lastname: Pierro
authors.forename: Pascale V
authors.initials: PV
authors.lastname: Guillot
authors.forename: Adrian J
authors.initials: AJ
authors.lastname: Thrasher
authors.forename: Paolo
authors.initials: P
authors.lastname: De Coppi
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology*; D000818:Animals; D015925:Cryopreservation; D004314:Down Syndrome; D005260:Female; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D051379:Mice; D008954:Models, Biological; D011247:Pregnancy; D011296:Prenatal Diagnosis