title: Routine isolation and expansion late mid trimester amniotic fluid derived mesenchymal stem cells in a cohort of fetuses with congenital diaphragmatic hernia.
abstract: OBJECTIVE
  To assess the feasibility of routine isolation and expansion of amniotic fluid derived mesenchymal stem cells (AF-MSC) in fetuses diagnosed with isolated congenital diaphragmatic hernia (CDH).
  
  
  METHODS
  Redundant AF samples of fetuses with CDH and normal fetuses were obtained. Cell colonies were mechanically selected for each sample. Proliferation capacity was expressed as population doubling time (PDT). Cell lines were further characterized with flow cytometry, differentiation assays and qRT-PCR (OCT4 and NANOG). After cell labeling with LacZ in vivo tracking was performed after fetal tracheal injection in rabbits.
  
  
  RESULTS
  Fourteen consecutive CDH samples (median gestational age (GA) of 32.9 weeks; IQR: 27.8-34.3 weeks) and seven control samples (30 weeks; IQR: 28.9-34.4 weeks) were obtained. PDT was similar in both groups (45.4h±1.9 vs. 52.3h±3.4;NS). AF-MSCs expressed a typical mesenchymal CD marker profile. Clones could be differentiated in osteogenic, adipogenic and chrondrogenic lineages. Expression of multipotency markers was low in all cell lines. We confirmed the presence of injected cells inside the fetal lung three days after intratracheal injection.
  
  
  CONCLUSIONS
  Routine isolation and expansion of AF-MSCs in CDH is feasible and cell lines generated were comparable to those of control samples. AF-MSCs from affected fetuses could potentially be used in future stem cell therapy.
authors.affiliation: Organ Systems Cluster, Department of Development and Regeneration, Center for Surgical Technologies, KU Leuven, Leuven, Belgium.
authors.forename: Philip
authors.initials: P
authors.lastname: DeKoninck
authors.affiliation: Organ Systems Cluster, Department of Development and Regeneration, Center for Surgical Technologies, KU Leuven, Leuven, Belgium.
authors.forename: Jaan
authors.initials: J
authors.lastname: Toelen
authors.affiliation: Organ Systems Cluster, Department of Development and Regeneration, Center for Surgical Technologies, KU Leuven, Leuven, Belgium.
authors.forename: Silvia
authors.initials: S
authors.lastname: Zia
authors.affiliation: Organ Systems Cluster, Department of Development and Regeneration, Center for Surgical Technologies, KU Leuven, Leuven, Belgium.
authors.forename: Maarten
authors.initials: M
authors.lastname: Albersen
authors.affiliation: Laboratory of Tissue Homeostasis and Disease, Skeletal Biology and Engineering Research Center, Development and Regeneration, KU Leuven, Leuven, Belgium.
authors.forename: Rik
authors.initials: R
authors.lastname: Lories
authors.affiliation: Department of Paediatric Surgery, Institute of Child Health & Great Ormond St. Hospital,London, United Kingdom.
authors.forename: Paolo De
authors.initials: PD
authors.lastname: Coppi
authors.affiliation: Organ Systems Cluster, Department of Development and Regeneration, Center for Surgical Technologies, KU Leuven, Leuven, Belgium. Electronic address: jan.deprest@uzleuven.be.
authors.forename: Jan
authors.initials: J
authors.lastname: Deprest
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology*; D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D002469:Cell Separation; D005260:Female; D065630:Hernias, Diaphragmatic, Congenital / Q000473:pathology*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D011247:Pregnancy; D011817:Rabbits
keywords: Amniotic fluid; Cell therapy; Congenital diaphragmatic hernia; Mesenchymal stem cells