The invention discloses a separating method and culturing method for hepatic stellate cells of a Mongolian gerbil. The separating method includes the steps that 1, the livers of the Mongolian gerbil are moved into a combined enzyme solution to be subjected to perfused digestion, then liver capsules and Glisson pedicles are removed, the livers are smashed to be pasty, the paste is moved into a combined enzyme solution, the mixture is stirred for digestion, and a stirred digestion product is obtained; 2, a first Hank's solution is added, the mixture is filtered, liver tissues are obtained, then a second Hank's solution is added, primary centrifuging is carried out, sediment is obtained, the sediment is suspended again through a third Hank's solution, then a Nycodenz solution is added, the mixture is mixed to be even, centrifuging is carried out again, and a turbid band, namely, the separated hepatic stellate cells is obtained. The separating and culturing method includes the steps of separating and culturing. By means of the methods, the yield and the survival rate of the hepatic stellate cells can be increased, the purity of the HSCs can be guaranteed, and a foundation is laid for further researching of deposition of cell matrixes and forming of fibrosis of the hepatic stellate cells after the livers are injured.