title: Molecular signature of amniotic fluid derived stem cells in the fetal sheep model of myelomeningocele.
abstract: Abnormal cord development results in spinal cord damage responsible for myelomeningocele (MMC). Amniotic fluid-derived stem cells (AFSCs) have emerged as a potential candidate for applications in regenerative medicine. However, their differentiation potential is largely unknown as well as the molecular signaling orchestrating the accurate spinal cord development. Fetal lambs underwent surgical creation of neural tube defect and its subsequent repair. AFSCs were isolated, cultured and characterized at the 12th (induction of MMC), 16th (repair of malformation), and 20th week of gestation (delivery). After performing open hysterectomy, AF collections on fetuses with sham procedures at the same time points as the MMC creation group have been used as controls. Cytological analyses with the colony forming unit assay, XTT and alkaline-phosphatase staining, qRT-PCR gene expression analyses (normalized with aged match controls) and NMR metabolomics profiling were performed. Here we show for the first time the metabolomics and molecular signature variation in AFSCs isolated in the sheep model of MMC, which may be used as diagnostic tools for the in utero identification of the neural tube damage. Intriguingly, PAX3 gene involved in the murine model for spina bifida is modulated in AFSCs reaching the peak of expression at 16 weeks of gestation, 4 weeks after the intervention. Our data strongly suggest that AFSCs reorganize their differentiation commitment in order to generate PAX3-expressing progenitors to counteract the MMC induced in the sheep model. The gene expression signature of AFSCs highlights the plasticity of these cells reflecting possible alterations of embryonic development.
authors.affiliation: Department of Public Health, Experimental and Forensic Medicine, Division of Human Anatomy, University of Pavia, Pavia, Italy.
authors.forename: Gabriele
authors.initials: G
authors.lastname: Ceccarelli
authors.affiliation: Department of Public Health, Experimental and Forensic Medicine, Division of Human Anatomy, University of Pavia, Pavia, Italy; Department of Development and Reproduction, Stem Cell Institute, Translational Cardiomyology Laboratory, KU Leuven, Leuven, Belgium.
authors.forename: Enrico
authors.initials: E
authors.lastname: Pozzo
authors.affiliation: Department of Maternal and Children's Health, Paediatric Surgery Unit, Fondazione IRCCS Policlinico San Matteo and University of Pavia, Pavia, Italy.
authors.forename: Federico
authors.initials: F
authors.lastname: Scorletti
authors.affiliation: Department of Public Health, Experimental and Forensic Medicine, Division of Human Anatomy, University of Pavia, Pavia, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Benedetti
authors.affiliation: Department of Public Health, Experimental and Forensic Medicine, Division of Human Anatomy, University of Pavia, Pavia, Italy.
authors.forename: Gabriella
authors.initials: G
authors.lastname: Cusella
authors.affiliation: Department of Pathology and Immunology, Faculty of Medicine, University of Geneva, Geneva, Switzerland.
authors.forename: Flavio Lorenzo
authors.initials: FL
authors.lastname: Ronzoni
authors.affiliation: Department of Development and Reproduction, Stem Cell Institute, Translational Cardiomyology Laboratory, KU Leuven, Leuven, Belgium.
authors.forename: Vardine
authors.initials: V
authors.lastname: Sahakyan
authors.affiliation: Department of Maternal and Children's Health, Paediatric Surgery Unit, Fondazione IRCCS Policlinico San Matteo and University of Pavia, Pavia, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Zambaiti
authors.affiliation: Department of Medical and Biological Sciences, University of Udine, Udine, Italy.
authors.forename: Maria Chiara
authors.initials: MC
authors.lastname: Mimmi
authors.affiliation: Department of Internal Medicine, University of Pavia and Department of Maternal and Children's Health, Paediatric Endocrinology and Diabetes Unit, Fondazione IRCCS Policlinico San Matteo, Pavia, Italy.
authors.forename: Valeria
authors.initials: V
authors.lastname: Calcaterra
authors.affiliation: Department Development and Regeneration, Organ System Cluster and Stem Cell Institute, Group Biomedical Sciences and Division Woman and Child, University Hospitals Leuven, KU Leuven, Leuven, Belgium.
authors.forename: Jan
authors.initials: J
authors.lastname: Deprest
authors.affiliation: Department of Public Health, Experimental and Forensic Medicine, Division of Human Anatomy, University of Pavia, Pavia, Italy; Department of Development and Reproduction, Stem Cell Institute, Translational Cardiomyology Laboratory, KU Leuven, Leuven, Belgium. Electronic address: sampa@unipv.it.
authors.forename: Maurilio
authors.initials: M
authors.lastname: Sampaolesi
authors.affiliation: Department of Maternal and Children's Health, Paediatric Surgery Unit, Fondazione IRCCS Policlinico San Matteo and University of Pavia, Pavia, Italy.
authors.forename: Gloria
authors.initials: G
authors.lastname: Pelizzo
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology* / Q000378:metabolism; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D005260:Female; D046128:Fetal Therapies / Q000379:methods*; D008591:Meningomyelocele / Q000628:therapy*; D055442:Metabolome; D055432:Metabolomics; D011247:Pregnancy; D012756:Sheep; D033581:Stem Cell Transplantation / Q000379:methods*; D013234:Stem Cells / Q000378:metabolism*
chemical_list: D015415:Biomarkers
keywords: Amniotic fluid derived stem cells; Gene signature; Myelomeningocele; Sheep model