title: Enhanced differentiation of human amniotic fluid-derived stem cells into insulin-producing cells in vitro.
abstract: OBJECTIVE
  To investigate the ability of human amniotic fluid stem cells (hAFSCs) to differentiate into insulin-producing cells.
  
  
  METHODS
  hAFSCs were induced to differentiate into pancreatic cells by a multistep protocol. The expressions of pancreas-related genes and proteins, including pancreatic and duodenal homeobox-1, insulin, and glucose transporter 2, were detected by polymerase chain reaction and immunofluorescence. Insulin secreted from differentiated cells was tested by enzyme-linked immunosorbent assay.
  
  
  RESULTS
  hAFSCs were successfully isolated from amniotic fluid that expressed the pluripotent markers of embryonic stem cells, such as Oct3/4, and mesenchymal stem cells, such as integrin β-1 and ecto-5'-nucleotidase. Here, we first obtained the hAFSCs that expressed pluripotent marker stage-specific embryonic antigen 1. Real-time polymerase chain reaction analysis showed that pancreatic and duodenal homeobox-1, paired box gene 4 and paired box gene 6 were expressed in the early phase of induction, and then stably expressed in the differentiated cells. The pancreas-related genes, such as insulin, glucokinase, glucose transporter 2 and Nkx6.1, were expressed in the differentiated cells. Immunofluorescence showed that these differentiated cells co-expressed insulin, C-peptide, and pancreatic and duodenal homeobox-1. Insulin was released in response to glucose stimulation in a manner similar to that of adult human islets.
  
  
  CONCLUSIONS
  The present study showed that hAFSCs, under selective culture conditions, could differentiate into islet-like insulin-producing cells, which might be used as a potential source for transplantation in patients with type 1 diabetes mellitus.
authors.affiliation: Department of Central Laboratory, China-Japan Union Hospital of Jilin University, Changchun, China.
authors.forename: Xu-Peng
authors.initials: XP
authors.lastname: Mu
authors.affiliation: College of Pharmacy, Jilin University, Changchun, China.
authors.forename: Li-Qun
authors.initials: LQ
authors.lastname: Ren
authors.affiliation: College of Pharmacy, Jilin University, Changchun, China.
authors.forename: Hao-Wei
authors.initials: HW
authors.lastname: Yan
authors.affiliation: College of Pharmacy, Jilin University, Changchun, China.
authors.forename: Xin-Min
authors.initials: XM
authors.lastname: Zhang
authors.affiliation: The Second Affiliated Hospital of Jilin University, Changchun, China.
authors.forename: Tian-Min
authors.initials: TM
authors.lastname: Xu
authors.affiliation: College of Pharmacy, Jilin University, Changchun, China.
authors.forename: An-Hui
authors.initials: AH
authors.lastname: Wei
authors.affiliation: Department of Central Laboratory, China-Japan Union Hospital of Jilin University, Changchun, China.
authors.forename: Jin-Lan
authors.initials: JL
authors.lastname: Jiang
mesh_terms: D050156:Adipogenesis; D000653:Amniotic Fluid / Q000166:cytology*; D002096:C-Peptide / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology*; D006801:Humans; D066298:In Vitro Techniques; D050417:Insulin-Secreting Cells / Q000502:physiology*; D007515:Islets of Langerhans / Q000502:physiology*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000502:physiology*; D055495:Neurogenesis
chemical_list: D002096:C-Peptide
keywords: Amniotic fluid-derived stem cells; Diabetes mellitus; Insulin-producing cells