title: [Establishment of new vessels model by implantation of VEGF165 in mouse].
abstract: OBJECTIVE
  To investigate the feasibility of establishing a murine new vessels model with Lentiviral vector (LVs) mediated human vascular endothelial growth factor-165 (pcDNA3.1/ VEGF165) gene.
  
  
  METHODS
  Lentivirus plasmid carrying a human VEGF165 was constructed and was used to transfect mouse's NIH/3T3 cells. The NIH/3T3 cells with high secretion of VEGF were injected into the skeletal muscle of mouse to establish a mouse new vessels model by implantation of vascular endothelium growth factor (VEGF) gene. The external secretion of VEGF was measured with ELISA. Histological examination was carried out after injection. The expression of CD31 was assessed with immunohistochemical method.
  
  
  RESULTS
  The lenti-VEGF165-EGFP was correctly constructed and confirmed by restriction endonuclease analysis, polymerase chain reaction and DNA sequencing analysis. Lentivirus plasmid carrying a human VEGF165 was constructed. lenti-VEGF165-EGFP was used to transfect mouse's NIH/ 3T3 cells, and human VEGF165 gene was assessed. NIH/3T3 cells mediated VEGF gene implantation can produce stable and effective mouse new vessels model. The external secretion of VEGF in peripheral blood was measured with ELISA. The legs became swollen in experimental group 14 d after injection. It found the cells expression of CD31 44 d after injection, and histological analysis showed the swollen tissue was composed of small new vessels.
  
  
  CONCLUSIONS
  NIH/3T3 cells mediated VEGF gene implantation can produce stable and effective mouse new vessels model.
authors.affiliation: Institute of Military Surgery, Xijing Hospital, Fourth Military University of Chinese PLA, Xi'an 710032, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Xie
authors.forename: Li
authors.initials: L
authors.lastname: Yang
authors.forename: Su-Yang
authors.initials: SY
authors.lastname: Tang
authors.forename: Yan
authors.initials: Y
authors.lastname: Zheng
authors.forename: Cheng-Gang
authors.initials: CG
authors.lastname: Yi
authors.forename: Wei
authors.initials: W
authors.lastname: Xia
authors.forename: Yong
authors.initials: Y
authors.lastname: Pan
authors.forename: Ge-Jia
authors.initials: GJ
authors.lastname: Ma
authors.forename: Yang
authors.initials: Y
authors.lastname: Yang
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D041681:NIH 3T3 Cells; D009389:Neovascularization, Pathologic; D010957:Plasmids; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics* / Q000378:metabolism
chemical_list: C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A