Objective To evaluate the autoclave sterilization effect on several kinds of animal cages and animal bedding, and provide the basis for safe sterilization operation. Methods Beddings and cages were sterilized with three different methods separately, and chemical indicator cards and biological indicator (Bacillus steatothermophilus) in (ATCC-7953) 3M^TM pressure-steam sterilization were used to indicate the sterilization effects. Results The beddings were not completely dried when it was sterilized in whole package. The sterilization quality was unstable while the beddings were autoclaved after divided into separate cages. The best sterilization effect and dry beddings were obtained if the whole package was autoclaved at first and then divided into separate cages and sterilized again. Conclusions The results of this study demonstrate that dry beddings and best sterilization effect can be achieved by whole package autoclaved at first followed by sterilization of the bedding material divided into separate cages.