An isolated animal organ perfusion device comprises a liquid storage assembly for storing a perfusion solution; the temperature control assembly is used for keeping the temperature of the perfusion solution in the conveying process; the oxygenation assembly is used for introducing oxygen into the conveyed perfusion solution to generate an oxygenation effect; the perfusion assembly comprises an outer-layer heat preservation subsystem and an inner-layer perfusion subsystem, the inner-layer perfusion subsystem comprises a perfusion chamber with a cover, a perfusion tube and a bile drainage tube are arranged on the cover, the perfusion chamber is divided into an upper layer and a lower layer by a perforated partition plate used for placing animal livers to be perfused, and the lower layer is connected back to the liquid storage assembly through a recovery pipeline. On the basis of realizing an animal perfusion technology, a controllable temperature environment and a perfusate oxygen concentration environment are provided. And more changeable and more perfect experiment conditions can be provided for related in-vitro animal organ tissue experiments. In the experiment process, the temperature probe, the oxygen concentration probe and the pH probe of the device can provide more experiment data.