Isolation of rat hepatic stellate cells by nonperfusion method and identification of isolated cells

Objective: To isolate rat hepatic stellate cells (HSCs) by nonperfusion method and identify the isolated cells. Methods: Rat liver cells were isolated by nonperfusion method combined with enzyme digestion method, from which HSCs were further isolated by density gradient centrifugation. The lipid droplets in cytoplasm of HSCs were determined by oil red staining, while the expressions of α-smooth muscle actin (α-SMA), desmin and neuroglia acid protein (GFAP) by immunohistochemical staining. Results: Rat HSCs were successfully isolated by nonperfusion method combined with enzyme digestion method. After oil red staining, red lipid droplets were found around the nucleus in HSCs purified by density gradient centrifugation. All the positive rates of α-SMA, desmin and GFAP were more than 90% in immunohistochemical staining. Conclusion: The nonperfusion isolating mode of rat HSC was established successfully, by which highly purified HSCs were obtained. The method was stable, simple, and worthy of popularization.