Generation of human induced pluripotent stem cells from extraembryonic tissues of fetuses affected by monogenic diseases

Introduction: The generation of human induced pluripotent stem cells (hiPSCs) derived from autologous extra-embryonic fetal source is an innovative personalized-regenerative technology, which 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 enables researchers to undertake studies for treating diseases or for future applications of in utero therapy. Methods: we use 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 deriving from prenatal diagnosis for several genetic disorder, such as Myotonic Dystrophy type 1 (DM1), β-Thalassemia (β-Thal), Lymphema-Distichiasis Syndrome (LDS), Spinal Muscular Atrophy (SMA), Cystic Fibrosis (CF), as well as from wild type (WT) fetal cells. Since cell type used for create iPSCs influence both the reprogramming process efficiency and the kinetics, we use chorionic villus (CVS) and amniotic fluid (AF) cells, demonstrating how they represent an ideal cell resource for efficient generation of hiPSCs. Results: The successful reprogramming of both CV and AF cells into hiPSCs is confirmed by specific morphological, molecular and immunocytochemical markers, and by their teratogenic potential when inoculated in vivo. We further demonstrate the stability of reprogrammed cells over 10 and more passages and their ability to differentiate into the three embryonic germ layers . Conclusions: These data suggest that hiPSCs-CVS/AF can be considered a valid cellular model to accomplish pathogenesis studies and therapeutic applications.