A new method for culturing vascular smooth muscle cells from the rabbit aorta / 中国药理学通报

ABSTRACT

Aim To develop a convenient and effective method to isolate and culture primary rabbit aortic vascular smooth muscle cells(VSMCs).Methods The thoracic aortas were removed by dissection under sterile conditions.Aortic smooth muscle cells were excised and cleaned of fat and connective tissue,and the isolated vessel media was cut into 1 mm3 pieces.The explants were digested with different concentrations of collagenase typeⅠ,and incubated at 37℃ for different time,then undispersed explants were placed onto a sterile 100-mm plastic tissue culture dish with growth medium.Results VSMCs could emigrate from the explants digested 6 h by collagenase typeⅠ(1.5 g?L-1)for 24 hours,the cells would passage for another 4~5 days.Confluency could be reached within 3~4 days after subculturing.VSMCs were identified by immunoreactivity with ?-actin and by the smooth muscle cell-specific,hills and valley-like morphology.Conclusion It was an effective method to culture primary VSMCs from the explants digested for 6h by collagenase type Ⅰ(1.5 g?L-1),which could shorten primary culture time.