title: Renal differentiation of amniotic fluid stem cells.
abstract: OBJECTIVE
  The role of stem cells in regenerative medicine is evolving rapidly. Here, we describe the application, for kidney regeneration, of a novel non-genetically modified stem cell, derived from human amniotic fluid. We show that these pluripotent cells can develop and differentiate into de novo kidney structures during organogenesis in vitro.
  
  
  METHODS
  Human amniotic fluid-derived stem cells (hAFSCs) were isolated from human male amniotic fluid obtained between 12 and 18 weeks gestation. Green fluorescent protein and Lac-Z-transfected hAFSCs were microinjected into murine embryonic kidneys (12.5-18 days gestation) and were maintained in a special co-culture system in vitro for 10 days. Techniques of live microscopy, histology, chromogenic in situ hybridization and reverse transcriptase polymerase chain reaction were used to characterize the hAFSCs during their integration and differentiation in concert with the growing organ.
  
  
  RESULTS
  Green fluorescent protein and Lac-Z-transfected hAFSCs demonstrated long-term viability in organ culture. Histological analysis of injected kidneys revealed that hAFSCs were capable of contributing to the development of primordial kidney structures including renal vesicle, C- and S-shaped bodies. Reverse transcriptase polymerase chain reaction confirmed expression of early kidney markers for: zona occludens-1, glial-derived neurotrophic factor and claudin.
  
  
  CONCLUSIONS
  Human amniotic fluid-derived stem cells may represent a potentially limitless source of ethically neutral, unmodified pluripotential cells for kidney regeneration.
authors.affiliation: Childrens Hospital Los Angeles, Saban Research Institute, Keck School of Medicine, University of Southern California, Los Angeles, CA 90027, USA.
authors.forename: L
authors.initials: L
authors.lastname: Perin
authors.forename: S
authors.initials: S
authors.lastname: Giuliani
authors.forename: D
authors.initials: D
authors.lastname: Jin
authors.forename: S
authors.initials: S
authors.lastname: Sedrakyan
authors.forename: G
authors.initials: G
authors.lastname: Carraro
authors.forename: R
authors.initials: R
authors.lastname: Habibian
authors.forename: D
authors.initials: D
authors.lastname: Warburton
authors.forename: A
authors.initials: A
authors.lastname: Atala
authors.forename: R E
authors.initials: RE
authors.lastname: De Filippo
mesh_terms: D000653:Amniotic Fluid / Q000166:cytology*; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D002465:Cell Movement; D002478:Cells, Cultured; D006801:Humans; D007668:Kidney / Q000166:cytology* / Q000196:embryology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008845:Microinjections; D013234:Stem Cells / Q000166:cytology*
chemical_list: D015415:Biomarkers