title: Second and third trimester amniotic fluid mesenchymal stem cells can repopulate a de-cellularized lung scaffold and express lung markers.
abstract: OBJECTIVE
  This study examined the potential of amniotic fluid mesenchymal stem cells (AF-MSCs) to generate lung precursor cells in vitro and on a xenologous three-dimensional de-cellularized lung scaffold.
  
  
  METHODS
  AF-MSCs were isolated from human amniotic fluid obtained from 17-37 weeks gestation. Lung differentiation was induced on Matrigel or on de-cellularized rat lungs intra-tracheally injected with AF-MSCs by culturing with a modification of small airway growth medium (mSAGM) lacking retinoic acid (RA) and triodothyronine (T3) with addition of fibroblast growth factor-10 (FGF10). Cells and scaffolds were characterized by immunofluorescence and RT-PCR for markers of viability, proliferation, and lung distal airway differentiation (TTF-1(+) and SPC(+)) in the absence of markers of brain (TuJ1(-)) and thyroid (Pax8(-)).
  
  
  RESULTS
  After culture in mSAGM on either Matrigel or lung scaffolds, there were TTF-1(+)/TuJ1(-)/Pax8(-) cells, indicating a lung precursor phenotype. In addition, SPC(+) cells also evolved suggesting a more mature lung phenotype.
  
  
  CONCLUSIONS
  We demonstrate that mid- to late-trimester AF-MSCs can be induced to develop into lung precursor cells when cultured on the appropriate extracellular matrix (ECM), making them a viable source for use in cell therapy or development of an ex vivo tissue engineered lung.
authors.affiliation: Department of Vascular Biology, University of Connecticut Health Center, 263 Farmington Avenue MC3501, Farmington, CT 06030.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Vadasz
authors.affiliation: Department of Vascular Biology, University of Connecticut Health Center, 263 Farmington Avenue MC3501, Farmington, CT 06030.
authors.forename: Todd
authors.initials: T
authors.lastname: Jensen
authors.affiliation: Department of Vascular Biology, University of Connecticut Health Center, 263 Farmington Avenue MC3501, Farmington, CT 06030.
authors.forename: Camilo
authors.initials: C
authors.lastname: Moncada
authors.affiliation: Department of Vascular Biology, University of Connecticut Health Center, 263 Farmington Avenue MC3501, Farmington, CT 06030; Department of Surgery, Connecticut Children's Medical Center, 282 Washington Street, Hartford, CT 06106.
authors.forename: Eric
authors.initials: E
authors.lastname: Girard
authors.affiliation: Department of Surgery, Connecticut Children's Medical Center, 282 Washington Street, Hartford, CT 06106.
authors.forename: Fan
authors.initials: F
authors.lastname: Zhang
authors.affiliation: Department of Vascular Biology, University of Connecticut Health Center, 263 Farmington Avenue MC3501, Farmington, CT 06030.
authors.forename: Alex
authors.initials: A
authors.lastname: Blanchette
authors.affiliation: Department of Vascular Biology, University of Connecticut Health Center, 263 Farmington Avenue MC3501, Farmington, CT 06030; Department of Surgery, Connecticut Children's Medical Center, 282 Washington Street, Hartford, CT 06106. Electronic address: cfinck@connecticutchildrens.org.
authors.forename: Christine
authors.initials: C
authors.lastname: Finck
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology*; D000818:Animals; D015415:Biomarkers; D002454:Cell Differentiation; D003094:Collagen; D004338:Drug Combinations; D005109:Extracellular Matrix; D005260:Female; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D007797:Laminin; D008168:Lung / Q000196:embryology; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D038081:Organogenesis; D010641:Phenotype; D011247:Pregnancy; D011262:Pregnancy Trimester, Second; D011263:Pregnancy Trimester, Third; D011509:Proteoglycans; D051381:Rats; D054457:Tissue Scaffolds
chemical_list: D015415:Biomarkers; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen
keywords: Amniotic fluid mesenchymal stem cells; De-cellularized lung; SPC; TTF-1