title: Differentiation of iPSCs into insulin-producing cells via adenoviral transfection of PDX-1, NeuroD1 and MafA.
abstract: OBJECTIVE
  The aim of this study was to evaluate the effect of PDX-1 (pancreatic and duodenal homeobox-1), NeuroD1 (neurogenic differentiation-1) and MafA (V-maf musculoaponeurotic fibrosarcoma oncogene homolog A) in the differentiation of induced pluripotent stem cells (iPSCs) into insulin-producing cells and to explore this new approach of cell transplantation therapy for type 1 diabetes in mice.
  
  
  METHODS
  iPSCs were infected with adenovirus (Ad-Mouse PDX-1-IRES-GFP, Ad-Mouse NeuroD1-IRES-GFP and Ad-Mouse Mafa-IRES-GFP) and then differentiated into insulin-producing cells in vitro. RT-PCR was applied to detect insulin gene expression, immunofluorescence to identify insulin protein, and mouse insulin enzyme-linked immunosorbent assay (ELISA) was used to evaluate the amount of insulin at different concentration of glucose. Insulin-producing cells were transplanted into the liver parenchyma of diabetic mice. Immunohistochemistry, intraperitoneal glucose tolerance test (IPGTT) and fasting blood glucose (FBG) were performed to assess the function of insulin-producing cells.
  
  
  RESULTS
  Insulin biosynthesis and secretion were induced in iPSCs and insulin-producing cells were responsive to glucose in a dose-dependent manner. Gene expression of the three-gene-modified embryoid bodies (EBs) was similar to the mouse pancreatic β cell line MIN6. Transplantation of insulin-producing cells into type I diabetic mice resulted in hyperglycemia reversal.
  
  
  CONCLUSIONS
  The insulin-producing cells we obtained from three-gene-modified EBs may be used as seed cells for tissue engineering and may represent a cell replacement strategy for the production of β cells for the treatment of type 1 diabetes.
authors.affiliation: Department of Surgical Comprehensive Laboratory, Affiliated Hospital of Nantong University, Nantong University, 226001 Nantong, PR China.
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
authors.affiliation: Department of Surgical Comprehensive Laboratory, Affiliated Hospital of Nantong University, Nantong University, 226001 Nantong, PR China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Nantong University, 226001 Nantong, PR China.
authors.forename: Qingsong
authors.initials: Q
authors.lastname: Guo
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Nantong University, 226001 Nantong, PR China.
authors.forename: Xiangjun
authors.initials: X
authors.lastname: Fan
authors.affiliation: Department of Surgical Comprehensive Laboratory, Affiliated Hospital of Nantong University, Nantong University, 226001 Nantong, PR China.
authors.forename: Yuhua
authors.initials: Y
authors.lastname: Lu
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Nantong University, 226001 Nantong, PR China.
authors.forename: Shajun
authors.initials: S
authors.lastname: Zhu
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Nantong University, 226001 Nantong, PR China.
authors.forename: Yao
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Surgical Comprehensive Laboratory, Affiliated Hospital of Nantong University, Nantong University, 226001 Nantong, PR China.
authors.forename: Xiangkun
authors.initials: X
authors.lastname: Bo
authors.affiliation: Department of Surgical Comprehensive Laboratory, Affiliated Hospital of Nantong University, Nantong University, 226001 Nantong, PR China.
authors.forename: Xu
authors.initials: X
authors.lastname: Chang
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Nantong University, 226001 Nantong, PR China.
authors.forename: Mingyan
authors.initials: M
authors.lastname: Zhu
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Nantong University, 226001 Nantong, PR China. Electronic address: wzw0417012168@163.com.
authors.forename: Zhiwei
authors.initials: Z
authors.lastname: Wang
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics* / Q000378:metabolism; D001786:Blood Glucose / Q000378:metabolism; D015153:Blotting, Western; D002454:Cell Differentiation / Q000502:physiology; D003922:Diabetes Mellitus, Type 1 / Q000378:metabolism / Q000473:pathology; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000166:cytology* / Q000378:metabolism; D051473:Maf Transcription Factors, Large / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D001786:Blood Glucose; D018398:Homeodomain Proteins; D007328:Insulin; D051473:Maf Transcription Factors, Large; D012333:RNA, Messenger; D015534:Trans-Activators; C485160:Mafa protein, mouse; C501008:Neurod1 protein, mouse; C084010:pancreatic and duodenal homeobox 1 protein
keywords: Insulin; Transgenes; Transplantation; iPSCs; β Cell