Aim To develop a method of culturing vascular smooth muscle cells (VSMC) from isolated rat thoracic aorta and compare their growth with that of aortic VSMC by other culture method. Methods The vascular media of rat thoracic aorta was isolated from male Wistar rats in sterile condition. Mince the media, and suspend the media in 10 mL 0.25% trypsin for about 3.5 h at 37℃. After terminating the digestion, centrifuge the solution at 100 g for 5 min, and seed the cell into a 25 cm2 vented cap flask. Results Thoracic aorta smooth muscle cell (SMC) were successfully passaged more than 20 times, without noticeable changes in morphology, growth characteristics, and smooth muscle α-actin expression. Conclusions Compared with the currently employed methods, our technique has the advantages of good reproducibility, high yield of pure SMC and quick growth of primary culture.