Objective To find a more appropriate method of adipogenic induction for adipose stem cells derived from humans and rats by comparing their adipogenic differentiation ability using two kinds of adipogenic culture media. Methods The human adipose stem cells (hADSCs) were extracted from lipoaspirates of 3 donors in the clinic, and the rat adipose stem cells (rADSCs) were obtained from adipose tissues of 6-week old rats. The cells were plated into two 6-well plates, and were divided into 3 groups: the negative control group, adipogenic induction group I and group II, respectively, two wells in each group. The control medium, adipogenic induction medium I and medium II were added into the cells of different groups, respectively. After induction for 10 days, the adipogenic differentiation ability of cells was assessed under microscope with oil red O staining and detecting the optical density value of 490 nm (A 490 nm) to compare the lipid droplet formation in the cells. Results The hADSCs and rADSCs showed a fibroblast-like growth. Positive oil red O staining cells showed orange lipid in the cells of adipogenic induction group from the third day of culture. The amount of lipid in cells induced by adipogenic induction medium II was higher than that in cells induced by adipogenic induction medium I, the A490 nm optical density of hADSCs induced by adipogenic induction medium II was significantly higher than that induced by adipogenic induction medium I (P<0.01), but there was no significant difference between the rADSCs induced by adipogenic induction medium I and II (P>0.05). Conclusions Both hADSCs and rADSCs can be induced to adipogenic differentiation using the two kinds of induction media, however, the induction of adipogenic differentiation ability of the adipogenic induction medium II is stronger than that of the adipogenic induction medium I.