Collagen has been widely used in biomedical field, such as scaffolds for tissue engineering. However, the rapidly biodegradation and weak mechanical strength of collagen limited its application. In this study, an insoluble collagen extracted from cattle hide was designed as scaffold to act as a three-dimensional substrate for tissue engineering. The received insoluble collagen sponge was analyzed by scanning electron microscopy, infrared spectroscopy, mechanical testing and thermogravimetric-differential thermal analysis. In addition, the degradation was performed in vitro using collagenase. The results showed that the insoluble collagen had the same triple helical domain as acid-soluble collagen, while the compression strength was greatly improved and the degradation rate was reduced. The insoluble collagen sponge with good stability should be promising in tissue engineering scaffold applications.