title: In vitro differentiation into insulin-producing β-cells of stem cells isolated from human amniotic fluid and dental pulp.
abstract: OBJECTIVE
  To investigate the ability of human amniotic fluid stem cells and human dental pulp stem cells to differentiate into insulin-producing cells.
  
  
  METHODS
  Human amniotic fluid stem cells and human dental pulp stem cells were induced to differentiate into pancreatic β-cells by a multistep protocol. Islet-like structures were assessed in differentiated human amniotic fluid stem cells and human dental pulp stem cells after 21 days of culture by dithizone staining. Pancreatic and duodenal homebox-1, insulin and Glut-2 expression were detected by immunofluorescence and confocal microscopy. Insulin secreted from differentiated cells was tested with SELDI-TOF MS and by enzyme-linked immunosorbent assay.
  
  
  RESULTS
  Human amniotic fluid stem cells and human dental pulp stem cells, after 7 days of differentiation started to form islet-like structures that became evident after 14 days of induction. SELDI-TOF MS analysis, revealed the presence of insulin in the media of differentiated cells at day 14, further confirmed by enzyme-linked immunosorbent assay after 7, 14 and 21 days. Both stem cell types expressed, after differentiation, pancreatic and duodenal homebox-1, insulin and Glut-2 and were positively stained by dithizone. Either the cytosol to nucleus translocation of pancreatic and duodenal homebox-1, either the expression of insulin, are regulated by glucose concentration changes. Day 21 islet-like structures derived from both human amniotic fluid stem cells and human dental pulp stem cell release insulin in a glucose-dependent manner.
  
  
  CONCLUSIONS
  The present study demonstrates the ability of human amniotic fluid stem cells and human dental pulp stem cell to differentiate into insulin-producing cells, offering a non-pancreatic, low-invasive source of cells for islet regeneration.
authors: Department of Surgery, Medicine, Dentistry and Morphological Sciences with Interest in Transplant, Oncology and Regenerative Medicine, University of Modena and Reggio Emilia, Modena, Italy. gianluca.carnevale@unimore.it
authors: Gianluca
authors: G
authors: Carnevale
authors: Massimo
authors: M
authors: Riccio
authors: Alessandra
authors: A
authors: Pisciotta
authors: Francesca
authors: F
authors: Beretti
authors: Tullia
authors: T
authors: Maraldi
authors: Manuela
authors: M
authors: Zavatti
authors: Gian Maria
authors: GM
authors: Cavallini
authors: Giovanni Battista
authors: GB
authors: La Sala
authors: Adriano
authors: A
authors: Ferrari
authors: Anto
authors: A
authors: De Pol
mesh_terms: D000653:Amniotic Fluid / Q000378:metabolism*; D002454:Cell Differentiation; D002478:Cells, Cultured; D003782:Dental Pulp / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D066298:In Vitro Techniques; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000378:metabolism*; D013234:Stem Cells / Q000378:metabolism*
chemical_list: D007328:Insulin
keywords: Diabetes; Insulin; hAFSCs; hDPSCs; β-Cells