Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (11): 1919-1922.doi: 10.3969/j.issn.1673-8225.2010.11.005

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Culture and identification of rabbit aortic endothelial cells: Modified isolation and culture of endothelial cells

Gao Hang, Guan Chun-yan   

  1. Department of Cardiology, First Affiliated Hospital of Liaoning Medical University, Jinzhou  121001, Liaoning Province, China
  • Online:2010-03-12 Published:2010-03-12
  • About author:Gao Hang, Master, Associate professor, Department of Cardiology, First Affiliated Hospital of Liaoning Medical University, Jinzhou 121001, Liaoning Province, China Shikongzhuren521@163.com

Abstract:

BACKGROUND: Although the methods of mechanical scrapping, tissue block transplantation, and enzyme digestion to culture endothelial cells have been quite mature, it has been constantly updated.
OBJECTIVE: To explore the culture and identification of rabbit aortic endothelial cells.
METHODS: Aorta was derived from one-week-old New Zealand rabbit. Following removing the adventitia, the endothelium was digested in the mixture of 2 g/L type Ⅰ collagen, 2 g/L type Ⅱ collagen, 2 g/L type Ⅳ collagen, and 2 g/L type Ⅴ collagen (1:1:1:1) for 20 minutes, and the digestion was stopped with culture medium (1:1). Cells in the endothelium were lightly scraped to make cell suspension, which was centrifuged to precipitate cells with DMEM culture medium containing 20% fetal bovine serum, 1 μg/L VEGF, 2 μg/L bFGF, and 6 U/L gentamicin. The cells were then blown into single cells and the culture liquid was changed every 48 hours. The cells were passaged according to the ratio of 1:2. Inverted phase contrast microscope was used to observe cell culture; immunohistochemistry and immunofluorescence were used to determine Ⅷ-related antigen; electron microscope was used to detect Weibel-Paladed body.
RESULTS AND CONCLUSION: VIII-related antigen and electron microscopy confirmed that the primary and 5-passage endothelial cells were successfully cultured, suggesting that rabbit aortic endothelial cells were cultured into cell line. VIII-related antigen and electron microscopy determined that a combination with W-P body was a good method to determine endothelial cells.

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