title: [Isolation, culture and identification of human umbilical vein endothelial cells].
abstract: OBJECTIVE
  To establish a simple, reliable and efficient isolation and culture method of human umbilical vein endothelial cells (HUVECs) in vitro.
  
  
  METHODS
  Type 2 collagenase was used to digest umbilical cord and separate HUVECs. The cells were cultured in the endothelial cell culture medium (ECM). The cell morphology was observed under an inverted phase-contrast microscope. Immunofluorescence technique was applied to detect the expression of von Willebrand factor (vWF). Cell purity was determined by detecting CD31 level on cell surface with flow cytometry. Tube formation assay was used to test the function of the endothelial cells after cryopreservation in vitro.
  
  
  RESULTS
  HUVECs successfully isolated were proved with high purity and good activity. HUVECs of primary generation could merge into a single layer one week after isolation. The cells showed a typical cobblestone-like arrangement. Immunofluorescence technique validated that the cells could widely express vWF and the expression frequency of CD31 was 93.1%. The cells were still capable of forming the lumen structure after cryopreservation, indicating that the standardized cryopreservation method could well maintain the cell function.
  
  
  CONCLUSIONS
  This is a simple, reliable and efficient method of isolating and culturing HUVECs in vitro.
authors.affiliation: Xinjiang Key Laboratory of Cardiovascular Disease, Clinical Medical Research Institute, Urumqi 830054, China.
authors.forename: Xiaocui
authors.initials: X
authors.lastname: Chen
authors.affiliation: Xinjiang Key Laboratory of Cardiovascular Disease, Clinical Medical Research Institute, Urumqi 830054, China.
authors.forename: Bangdang
authors.initials: B
authors.lastname: Chen
authors.affiliation: Xinjiang Key Laboratory of Cardiovascular Disease, Clinical Medical Research Institute, Urumqi 830054, China.
authors.forename: Yining
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Xinjiang Key Laboratory of Cardiovascular Disease, Clinical Medical Research Institute, Urumqi 830054, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Xinjiang Key Laboratory of Cardiovascular Disease, Clinical Medical Research Institute, Urumqi 830054, China.
authors.forename: Fen
authors.initials: F
authors.lastname: Liu
authors.affiliation: Xinjiang Key Laboratory of Cardiovascular Disease, Clinical Medical Research Institute, Urumqi 830054, China.
authors.forename: Mintao
authors.initials: M
authors.lastname: Gai
authors.affiliation: Xinjiang Key Laboratory of Cardiovascular Disease, Clinical Medical Research Institute, Urumqi 830054, China.
authors.forename: Qingjie
authors.initials: Q
authors.lastname: Chen
authors.affiliation: Xinjiang Key Laboratory of Cardiovascular Disease, Clinical Medical Research Institute, Heart Center, First Affiliated Hospital, Xinjiang Medical University, Urumqi 830054, China.
authors.forename: Yitong
authors.initials: Y
authors.lastname: Ma
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation / Q000502:physiology; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D015925:Cryopreservation; D005260:Female; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D006801:Humans; D007231:Infant, Newborn; D020784:Matrix Metalloproteinase 8 / Q000378:metabolism; D008858:Microscopy, Phase-Contrast; D018919:Neovascularization, Physiologic / Q000502:physiology; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D015203:Reproducibility of Results; D013997:Time Factors; D014470:Umbilical Cord / Q000166:cytology* / Q000378:metabolism; D014841:von Willebrand Factor / Q000378:metabolism
chemical_list: D019408:Platelet Endothelial Cell Adhesion Molecule-1; D014841:von Willebrand Factor; D020784:Matrix Metalloproteinase 8