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: Paola Spitalieri, Rosa V Talarico, Annalisa Botta, Michela Murdocca, Maria Rosaria D'Apice, Augusto Orlandi, Emiliano Giardina, Massimo Santoro, Francesco Brancati, Giuseppe Novelli, Federica Sangiuolo
• Journal: Cellular reprogramming (Cell Reprogram) Vol. 17 Issue 4 Pg. 275-87 (Aug 2015) ISSN: 2152-4998 [Electronic] United States
• PMID: 26474030 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• KLF4 protein, human
• Klf4 protein, mouse
• Kruppel-Like Factor 4
• Kruppel-Like Transcription Factors
• MYC protein, human
• Octamer Transcription Factor-3
• POU5F1 protein, human
• Proto-Oncogene Proteins c-myc
• SOX2 protein, human
• SOXB1 Transcription Factors

Topics

• Animals
• Cells, Cultured
• Cellular Reprogramming
• Chorionic Villi Sampling
• Cystic Fibrosis (embryology)
• Eyelashes (abnormalities, embryology)
• Female
• Fetus (cytology, physiology)
• Genetic Vectors
• Humans
• Induced Pluripotent Stem Cells (metabolism)
• Kruppel-Like Factor 4
• Kruppel-Like Transcription Factors (genetics)
• Lentivirus (genetics)
• Lymphedema (embryology)
• Mice
• Muscular Atrophy, Spinal (embryology)
• Myotonic Dystrophy (embryology)
• Octamer Transcription Factor-3 (genetics)
• Pregnancy
• Proto-Oncogene Proteins c-myc (genetics)
• SOXB1 Transcription Factors (genetics)
• Transgenes
• beta-Thalassemia (embryology)