Objective：To establish a simple collagenase perfusion method through abdominal aorta cannulation to isolate rat pancreatic stellate cell and demonstrate the procedures of its culture and characterization. Methods：Via a cannula iserted into adominal aorta, Gey’s balanced salt solution containing 0.025% collagenase P was perfused into rat normal pancreas. After digestion with collagenase P and protease, Nycodenz density gradient centrifugation was performed to isolate PSC. PSC was identified by its morphology, cytoplasmic lipid droplets and immunocytochemical staining for desmin, GFAP and α-SMA. Results：The production,viability and purity of isolated PSC were（15.3 ± 4.6） × 103／g body weight, （95.0 ± 3.5）%, and ﹥80％ respectively. After 24 hours in culture, most PSC attached to culture dishes, and showed angular or stellate appearance. PSC became positive for desmin and GFAP after 48 hours in culture but did not express α-SMA until 96 hours in culture. Conclusion：The collagenase perfusion method through abdominal aorta cannulation could be used for PSC isolation, with high production, viability and purity, therefore, it could to meet the need of in vitro experiments.