Lots of research has indicated that materials contain Arg-Gly-Asp (RGD) sequence can promote cell attachment and proliferation on them. Although Antheraea pernyi silk fibroin is a natural structural protein which contains RGD sequence, there are few studies on this kind of protein materials, for the regeneration of Antheraea pernyi silk fibroin from silk fibers is complicated and it is hard to be processed. In this paper, we present a water-insoluble Antheraea pernyi/Bombyx mori silk fibroin blending scaffold. The regenerated Antheraea pernyi silk fibroin (RASF) solution was prepared by dissolving degummed silk fibers at 100℃ and dialyzing at 4℃. The regenerated Bombyx mori silk fibroin (RBSF) solution was prepared by dissolving degummed silk fibers at 60℃ and dialyzing at 20℃. Regenerated silk fibroin solution was concentrated to 6 wt% solution in 10 wt% PEG solution. Based on RASF and RBSF solution, RBSF porous scaffold and RASF/RBSF blending scaffolds with different ratios were prepared through treating 1-butanol/SF solution under freezing at -20℃. The volume ratio of 1-butanol to solution was 1:2. RASF porous scaffold was not hard enough to hold itself, therefore the maximum content of RASF in blending scaffold was 70 wt%. With increasing of RASF content, pore sizes of scaffolds decreased from 250 μm to 150 μm and compressive strengths decreased from 280 kPa to 108 kPa, while the thermal stabilities increased. FTIR results demonstrated that the molecular conformation of silk fibroin was proven to be β-sheet, β-turn and α-helix. The biocompatibilities of scaffolds were demonstrated with in vitro cell culture. The results showed that L929 fibroblast and MC3t3-E1 osteoblast adhered, proliferated and migrated well into the scaffolds. The speed of cell proliferation accelerated with the increase of RASF content. Obviously, these regenerated silk fibroin scaffolds with good bio-compatibility could be used in tissue engineering field further.