title: [Differentiation of polygene-modified bone marrow mesenchymal stem cells into insulin-producing cells].
abstract: OBJECTIVE
  To evaluate the effects of insulin gene transcription regulators PDX-1, NeuroD1 and MafA on the differentiation of bone marrow mesenchymal stem cells (mMSCs) into insulin-producing cells.
  
  
  METHODS
  Murine mMSCs were isolated, cultured and expanded. The base sequences of transcription factors PDX-1, NeuroD1 and MafA were obtained by total gene synthesis and the recombinant adenovirus vectors harboring target genes constructed and transfected into packaging cell line 293A. mMSCs were infected with adenovirus separately or together, and then differentiated in vitro into insulin-producing cells. Reverse transcription-polymerase chain reaction (RT-PCR) was utilized to detect insulin gene expression, immunofluorescence for identifying the presence of insulin protein and insulin enzyme-linked immunosorbent assay (ELISA) for evaluating the secretory volume of insulin.
  
  
  RESULTS
  The differentiation extent of mMSCs into β-cell was analyzed. The β-cell-specific transcriptional regulators and insulin gene were expressed in mMSCs after transfection. Immunofluorescent analyses revealed the activated expression of insulin in the cytoplasm of differentiated cells. A significant content of insulin was released in these cells in response to a certain concentrations of glucose stimulation. The insulin content of mMSCs infected with a combination of three transcription factors was significantly higher than that of the control group [(112.84 ± 9.67) mU/L vs (1.60 ± 0.22) mU/L, P < 0.05].
  
  
  CONCLUSIONS
  After modification by transcriptional factors PDX-1, NeuroD1 and MafA, mMSCs can secrete insulin through starting endogenous insulin gene transcription.
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Nantong 226001, China.
authors.forename: Qing-song
authors.initials: QS
authors.lastname: Guo
authors.forename: Ming-yan
authors.initials: MY
authors.lastname: Zhu
authors.forename: Xiang-jun
authors.initials: XJ
authors.lastname: Fan
authors.forename: Yu-hua
authors.initials: YH
authors.lastname: Lu
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
authors.forename: Zhi-wei
authors.initials: ZW
authors.lastname: Wang
mesh_terms: D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics; D001854:Bone Marrow Cells / Q000166:cytology*; D002454:Cell Differentiation; D015870:Gene Expression; D005822:Genetic Vectors; D018398:Homeodomain Proteins / Q000235:genetics; D050417:Insulin-Secreting Cells / Q000166:cytology*; D051473:Maf Transcription Factors, Large / Q000235:genetics; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051379:Mice; D009419:Nerve Tissue Proteins / Q000235:genetics; D030781:Organisms, Genetically Modified; D015534:Trans-Activators / Q000235:genetics
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D018398:Homeodomain Proteins; D051473:Maf Transcription Factors, Large; D009419:Nerve Tissue Proteins; D015534:Trans-Activators; C485160:Mafa protein, mouse; C084010:pancreatic and duodenal homeobox 1 protein; C093564:Neurogenic differentiation factor 1