Modified explant technique for the culture of human vascular smooth muscle cells

Abstract

Objective To establish an effective method of human vascular smooth muscle cells in vitro culture.Methods Medial layer of human aorta was separated under sterile conditions,then cut into small pieces (about 1 mm3).After pretreatment with 0.1％ type Ⅰ collagenase,the tissues were transferred into flask and cultured in DMEM containing 20％ fetal bovine serum,penicillin (100 U/ml) and streptomycin (100 g/L).The cell morphology was observed under the inverted microscopy.Smooth muscle cell specific protein (α-SMA) was identified by using immumofluorescence methods.Results Some smooth muscle cells started growing from explants in 4 to 5 days.One to 2 weeks later,the cells grew to confluence by migrated cells.Immumofluorescence staining showed positive expression of α-SMA (fluorescence intensity (++)-(+++)).Conclusion Modified explant technique by pretreating tissues with collagenase was an effective method to get vascular smooth muscle cells in vitro. Key words: Aorta; Vascular smooth muscle cells; Cell culture