title: Generation of Human Induced Pluripotent Stem Cells from Extraembryonic Tissues of Fetuses Affected by Monogenic Diseases.
abstract: The generation of human induced pluripotent stem cells (hiPSCs) derived from an autologous extraembryonic fetal source is an innovative personalized regenerative technology that can transform own-self cells into embryonic stem-like ones. These cells are regarded as a promising candidate for cell-based therapy, as well as an ideal target for disease modeling and drug discovery. Thus, hiPSCs enable researchers to undertake studies for treating diseases or for future applications of in utero therapy. We used a polycistronic lentiviral vector (hSTEMCCA-loxP) encoding OCT4, SOX2, KLF4, and cMYC genes and containing loxP sites, excisible by Cre recombinase, to reprogram patient-specific fetal cells derived from prenatal diagnosis for several genetic disorders, such as myotonic dystrophy type 1 (DM1), β-thalassemia (β-Thal), lymphedema-distichiasis syndrome (LDS), spinal muscular atrophy (SMA), cystic fibrosis (CF), as well as from wild-type (WT) fetal cells. Because cell types tested to create hiPSCs influence both the reprogramming process efficiency and the kinetics, we used chorionic villus (CV) and amniotic fluid (AF) cells, demonstrating how they represent an ideal cell resource for a more efficient generation of hiPSCs. The successful reprogramming of both CV and AF cells into hiPSCs was confirmed by specific morphological, molecular, and immunocytochemical markers and also by their teratogenic potential when inoculated in vivo. We further demonstrated the stability of reprogrammed cells over 10 and more passages and their capability to differentiate into the three embryonic germ layers, as well as into neural cells. These data suggest that hiPSCs-CV/AF can be considered a valid cellular model to accomplish pathogenesis studies and therapeutic applications.
authors.affiliation: 1 Department of Biomedicine and Prevention, Tor Vergata University of Rome , Rome, 00133, Italy .
authors.forename: Paola
authors.initials: P
authors.lastname: Spitalieri
authors.affiliation: 1 Department of Biomedicine and Prevention, Tor Vergata University of Rome , Rome, 00133, Italy .
authors.forename: Rosa V
authors.initials: RV
authors.lastname: Talarico
authors.affiliation: 1 Department of Biomedicine and Prevention, Tor Vergata University of Rome , Rome, 00133, Italy .
authors.forename: Annalisa
authors.initials: A
authors.lastname: Botta
authors.affiliation: 1 Department of Biomedicine and Prevention, Tor Vergata University of Rome , Rome, 00133, Italy .
authors.forename: Michela
authors.initials: M
authors.lastname: Murdocca
authors.affiliation: 2 Department of Laboratory Medicine, Policlinic of Tor Vergata , Rome, 00133, Italy .
authors.forename: Maria Rosaria
authors.initials: MR
authors.lastname: D'Apice
authors.affiliation: 1 Department of Biomedicine and Prevention, Tor Vergata University of Rome , Rome, 00133, Italy .
authors.forename: Augusto
authors.initials: A
authors.lastname: Orlandi
authors.affiliation: 1 Department of Biomedicine and Prevention, Tor Vergata University of Rome , Rome, 00133, Italy .|3 Molecular Genetics Laboratory UILDM , Santa Lucia Foundation, Rome, 00142, Italy .
authors.forename: Emiliano
authors.initials: E
authors.lastname: Giardina
authors.affiliation: 4 Don Carlo Gnocchi Foundation, ONLUS , Milan, 20148, Italy .
authors.forename: Massimo
authors.initials: M
authors.lastname: Santoro
authors.affiliation: 2 Department of Laboratory Medicine, Policlinic of Tor Vergata , Rome, 00133, Italy .
authors.forename: Francesco
authors.initials: F
authors.lastname: Brancati
authors.affiliation: 1 Department of Biomedicine and Prevention, Tor Vergata University of Rome , Rome, 00133, Italy .|2 Department of Laboratory Medicine, Policlinic of Tor Vergata , Rome, 00133, Italy .
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Novelli
authors.affiliation: 1 Department of Biomedicine and Prevention, Tor Vergata University of Rome , Rome, 00133, Italy .|2 Department of Laboratory Medicine, Policlinic of Tor Vergata , Rome, 00133, Italy .
authors.forename: Federica
authors.initials: F
authors.lastname: Sangiuolo
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D015193:Chorionic Villi Sampling; D003550:Cystic Fibrosis / Q000196:embryology; D005140:Eyelashes / Q000002:abnormalities / Q000196:embryology; D005260:Female; D005333:Fetus / Q000166:cytology* / Q000502:physiology; D005822:Genetic Vectors; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism*; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000235:genetics; D016086:Lentivirus / Q000235:genetics; D008209:Lymphedema / Q000196:embryology; D051379:Mice; D009134:Muscular Atrophy, Spinal / Q000196:embryology; D009223:Myotonic Dystrophy / Q000196:embryology; D050814:Octamer Transcription Factor-3 / Q000235:genetics; D011247:Pregnancy; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics; D055748:SOXB1 Transcription Factors / Q000235:genetics; D019076:Transgenes; D017086:beta-Thalassemia / Q000196:embryology
chemical_list: C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; C489427:MYC protein, human; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D016271:Proto-Oncogene Proteins c-myc; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors