title: [Isolation, culture, and identification of canine umbilical vein vascular endothelial cells].
abstract: OBJECTIVE
  To establish a simple and efficient method to isolate and culture the umbilical vein vascular endothelial cells in canine.
  
  
  METHODS
  Twelve umbilical cords [(13.0 +/- 1.5) cm in length] were taken from 12 newborn pups of Beagles. And then the vascular endothelial cells were isolated from these umbilical cords digested by 1% collagenase type I for 5, 7, and 10 minutes respectively (4 umbilical cords in each group). After cultured, the vascular endothelial cells were identified by morphology, immunofluorescence, and flow cytometry. And the growth curvature of umbilical vein vascular endothelial cells was detected by MTT assay.
  
  
  RESULTS
  Few vascular endothelial cells were collected at 5 and 10 minutes after digestion; many vascular endothelial cells were seen at 7 minutes, and became cobblestone with culture time, with a large nucleus; after passage, cell morphology had no obvious change. Fluorescence microscope results showed that positive von Willebrand factor (vWF) and CD31 cells were observed in most of cells. The flow cytometry test displayed that the positive cell rates of vWF and CD31 were 99.0% +/- 0.7% and 98.0% +/- 1.2%, respectively. The above results indicated that cultured cells were vascular endothelial cells. MTT assay showed that vascular endothelial cells proliferation increased significantly with culture time.
  
  
  CONCLUSIONS
  Enzyme digestion is a convenient method to isolate vascular endothelial cells from canine umbilical vein, and a large number of cells and high purity of cells can be obtained by the method.
authors.affiliation: Burns Institute, the First Affiliated Hospital of Chinese PLA General Hospital, Beijing, 100048, P.R.China.
authors.forename: Quan
authors.initials: Q
authors.lastname: Hu
authors.forename: Jiake
authors.initials: J
authors.lastname: Chai
authors.forename: Lingying
authors.initials: L
authors.lastname: Liu
authors.forename: Yusen
authors.initials: Y
authors.lastname: Hou
authors.forename: Yihe
authors.initials: Y
authors.lastname: Wang
authors.forename: Bailing
authors.initials: B
authors.lastname: Li
authors.forename: Hongming
authors.initials: H
authors.lastname: Yang
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D017364:Collagenases; D004285:Dogs; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology; D005260:Female; D005434:Flow Cytometry; D008856:Microscopy, Fluorescence / Q000379:methods; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D014470:Umbilical Cord / Q000166:cytology; D014471:Umbilical Veins / Q000166:cytology*; D014841:von Willebrand Factor / Q000378:metabolism
chemical_list: D019408:Platelet Endothelial Cell Adhesion Molecule-1; D014841:von Willebrand Factor; D017364:Collagenases