Objective To evaluate a new method for the isolation of rat pancreatic stellate cells (PSCs) and to investigate the influence of adipose derived stem cells (ADSCs) on PSCs in vitro.Methods Normal rat PSCs was isolated by collagenase and Optiprep density gradient centrifugation.The coculture system of ADSCs and PSCs was set up by transwell insert.The proliferation of PSCs was tested by CCK-8 test kit.Smoothmuscle α-actin (α-SMA) expression of PSCs were tested by Western blot.The apoptosis of HSCs was tested by flow cytometer.The cytokines in the culture solution were tested by ELISA kit.Results The quantity of PSCs was above 5 × 106 cells per rat.The purity and the viability of the cells were about 90-97 percent.After coculture for 72 h,the proliferation and activation of PSCs was inhibited by ADSCs (F =223.27,P ＜ 0.05 ; F =52.97,P ＜ 0.05) and the apoptosis of PSCs was promoted by ADSCs (F =43.62,P ＜ 0.05).more NGF and less TGF-β1 was secreted by ADSCs than by PSCs (NGF:14.68 ± 0.94 vs.8.31 ±0.86,t =4.67,P ＜0.05;TGF-β1:10.65 ±0.46 vs.70.47 ±0.99,t =21.72,P＜0.01).Conclusions ADSCs inhibit the proliferation and activation of PSCs by ADSCs secreting cytokines.