TY - JOUR
T1 - Trisomy 21 mid-trimester amniotic fluid induced pluripotent stem cells maintain genetic signatures during reprogramming
T2 - Implications for disease modeling and cryobanking
AU - Pipino, Caterina
AU - Mukherjee, Sayandip
AU - David, Anna L.
AU - Blundell, Michael P.
AU - Shaw, Steven W.
AU - Sung, Peggy
AU - Shangaris, Panicos
AU - Waters, Jonathan J.
AU - Ellershaw, Drew
AU - Cavazzana, Marina
AU - Mostoslavsky, Gustavo
AU - Pandolfi, Assunta
AU - Pierro, Agostino
AU - Guillot, Pascale V.
AU - Thrasher, Adrian J.
AU - De Coppi, Paolo
PY - 2014/9/22
Y1 - 2014/9/22
N2 - 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 72h). 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.
AB - 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 72h). 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.
UR - http://www.scopus.com/inward/record.url?scp=84907278141&partnerID=8YFLogxK
U2 - 10.1089/cell.2013.0091
DO - 10.1089/cell.2013.0091
M3 - Article
C2 - 25162836
AN - SCOPUS:84907278141
SN - 2152-4971
VL - 16
SP - 331
EP - 344
JO - Cellular Reprogramming
JF - Cellular Reprogramming
IS - 5
ER -