A new method for culturing endothelial cells from the rat aorta / 中国药理学通报

ABSTRACT

Aim To develop a convenient and effective method to isolate and culture primary rat aortic endothelial cells (RAECs). Methods The thoracic aortas were removed by dissection under sterile conditions. Aortas were turned over to expose the luminal surface, and the surfaces were digested with different concentrations of collagenase typeⅠ, incubated at 37℃ for different times, then, cut into pieces and placed luminal side down onto collagen-coated flask with growth medium. Results RAECs could emigrate from explants digested 1h by collagenase typeⅠ(2.0 g?L-1) for 24 h and cells would passage for another 4~5 days. Reached confluency within 3~4 d after subculturing. RAECs were identified by immunoreactivity with Factor-Ⅷ and by the endothelial cell-specific, cobblestone-like morphology. Conclusion It is an effective method to culture primary RAECs from explants digested for 1h by collagenase type Ⅰ(2.0 g?L-1),that can shorten primary culture time.