Abstract：Objective: To establish a simple and efficient method for primary culture of mouse aortic vascular smooth muscle cells (VSMCs) and calcification model in vitro. Methods: The primary VSMCs were harvested by modified tissue-piece inoculation and identified by immunocytochemistry. The established VSMCs were randomly divided into control group and calcification group. Calcification of cells was assayed by von kossa staining and colorimetry method. Results: VSMCs migrated from explants of mouse aorta tissue after 3-5 days of culture. After 7-10 days, VSMCs grew to form a fusing monolayer.Immunofluorescence staining with specific mAb against mouse α-actin demonstrated VSMCs were positive. The cell purity of the 2nd generation of VSMCs was over 95%. Compared with the control group, the calcium content and ALP activity in the calcification group were increased significantly (both P<0.05). Conclusion: The method of modified explant-culture successfully established a effective model for primary culture of VSMCs, of which calcification in vitro could be induced by β-glycerophosphate.
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2014, Vol. 24 
