Decellularization means and altering crosslinking approaches were two promising alternatives for glutaraldehyde fixation to biological tissues. Bovine jugular veins (BJVs) were decellularized by a multi-step detergent-enzymatic extraction method, then photooxidatively crosslinked. Gross and histological integrity of which was retained. Ultrastructures showed integrity of collagen fibrils and elastic fibers, and a basement membrane free luminal surface. Mechanical strength test and tissue protein extraction assay demonstrated their tissue stability. After being pre-coated with gelatin, collagen IV and fibronectin, cultured human umbilical vein endothelial cells were planted in the luminal surface of decellularized plus photooxidized BJV patches for seven days. Endothelial cells were denser in pre-coated patches than in uncoated controls. A rat subcutaneous implantation model revealed more resistance against in vivo degradation for further crosslinked BJV patches than decellularized patches at 12-week retrieval. Host cells were all layer repopulated for both. Histological examination and content assay demonstrated collagen and glycosaminoglycan components synthesis for decellularized plus photooxidized BJV patches. Decellularized and photooxidatively crosslinked BJV patches possess tissue integrity, excellent in vitro and in vivo tissue stability and repopulation patterns. Thus, they have potentials as tissue engineering scaffolds in future cardiovascular surgery.