Culture of vascalar smooth muscle cells from rat cerebral vessels by enzymatic dispersion method / 中国药理学通报

ABSTRACT

Aim To develop a method of culturing rat basilar artery smooth muscle cells. Methods Basilar arteries were immediately removed and cut into 0.2 mm rings. Then they were incubated in a medium containing the following: collagenase (typeⅡ, 0.5 g?L-1), elastase (0.15 g?L-1), hyaluronidase (type Ⅳ-S, 0.5 g?L-1), and deoxyribonuclease (typeⅠ, 0.1 g?L-1) for 1 hour at 37℃. After digestion, cells were cultured in DMEM/F12 with 20% fetal calf serum and then plated onto plastic tissue culture dishes. Results After 3 days of incubation, primary cultures of rat basilar artery smooth muscle cells began to stick to the wall of the incubation dishes. Cells were passaged after one week and their viability was 97%. The purity of fourth passage cells was 98%, which assessed by immunohistochemical staining and morphology. Conclusion This method is simple, fast and easily adaptable to obtaining high yield and purity of basilar artery smooth muscle cells.