Chinese Journal of Tissue Engineering Research ›› 2016, Vol. 20 ›› Issue (7): 1019-1024.doi: 10.3969/j.issn.2095-4344.2016.07.015

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Improved extraction of primary vascular endothelial cells from the rabbit aorta

Xiao Zi-chun, Tan Jin-hai, Zhou Yi   

  1. Department of Orthopedics, Zhongnan Hospital of Wuhan University, Wuhan 430071, Hubei Province, China
  • Received:2016-01-04 Online:2016-02-12 Published:2016-02-12
  • Contact: Tan Jin-hai, M.D., Chief physician, Professor, Master’s supervisor, Department of Orthopedics, Zhongnan Hospital of Wuhan University, Wuhan 430071, Hubei Province, China
  • About author:Xiao Zi-chun, Studying for master’s degree, Department of Orthopedics, Zhongnan Hospital of Wuhan University, Wuhan 430071, Hubei Province, China
  • Supported by:

    the Natural Science Foundation of Hubei Province, No. 2014CFA084

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Abstract:

BACKGROUND: Primary vascular endothelial cells are mostly harvested through aorta endothelial cell cultures and micro-artery endothelial cell cultures using enzyme digestion and tissue adhesion methods, and the quality and purity of harvested cells cannot meet the need for current scientific research.
OBJECTIVE: To investigate an improved extraction of primary vascular endothelial cells and the relevant identification method.
METHODS: A segment of rabbit aorta was cut to culture vascular endothelial cells using the improved extraction method in group A or using adhesion method in group B. In the group A, the vascular intima was striped out with microsurgical instruments, and digested enzymatically to acquire single primary cells followed by culture in endothelial cell culture medium. In the group B, the whole vascular intima was adhered to the culture dish that was incubated in a 5%CO2, 37 ℃ incubator for 1 hour. Cell pellets in the two groups were cultured in vitro. Cell morphology was observed using a microscope; immunohistochemical staining was used to detect CD31, VIII factor and Vimentin protein for identification of vascular endothelial cells.
RESULTS AND CONCLUSION: The purity and number of vascular endothelial cells extracted by the improved method were higher than those by the adhesion method. Immunohistochemical findings showed positive expression of CD31 and VIII factor, but negative expression of Vimentin. These findings indicate that the improved extraction method can obtain more vascular endothelial cells with higher purity, which is of strong operability and practicality.