Generation of disease-specific induced pluripotent stem cells from human fetal extra-embryonic tissues

      The generation of induced pluripotent stem cells (iPSCs) is a innovative personalized-regenerative technology, which can transform own-self cells into embryonic stem –like cells, which have regarded as a promising candidate for cell-based therapy, as well as an ideal target for disease modeling and drug testing and drug discovery, thus enabling researchers to undertake studies for treating diseases. The objective of the present study was to reprogramming patient-specific fetal cells deriving from prenatal diagnosis for several genetic disorder as Cystic Fibrosis (FC), Myotonic Dystrophy (DM1), β-Thalassemia (β-Thal), Spinal Muscolar Atrophy (SMA1), Lymphema-Distichiasis Syndrome (FOXC2) and healthy cells. The cell type used for create iPSCs can significantly influence the reprogramming efficiency and kinetics. Here, we show that amniotic fluid (LA) and chorionic villus sampling (CVS) represent an ideal cell resource for rapid and efficient generation of human iPSCs. The reprogramming were done using a polycistronic lentiviral vector (hSTEMCCA-loxP) encoding Oct4, Sox2, Klf4 and c-Myc genes necessary to cell reprogramming. Moreover loxP sites can be excised with Cre recombinase. Stem cells specific morphological, molecular and immunocytochemical markers (ALP; OCT4; SSEA4; TRA1-60;TRA1-81) confirmed the successful reprogramming. Additionally, we evaluated their ability to differentiate into the three embryonic germ layers (ecto, endo and mesoderm) by immunocytochemical characterization and their ability to in vivo form teratomas. To date, this represents the first example of iPS cells derived from a very early extra-embryonic fetal tissues like chorionic villi (hIPS-CVS). These data suggest that hIPS-CVS/LA can be considered a valid cell model to accomplish pathogenesis studies and possibly represent a valid tool for future therapeutic applications.
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