Decellularized tissues and organs from animal sources are widely used in regenerative medicine and tissue engineering. However, in tendon tissue engineering there is limitation not only in terms of tissue source -allografts and autografts- but also standardization of decellularization techniques. The goal of this study is to decellularize porcine Achilles tendon to be used as an off-the-shelf product for tendon reconstruction. We describe a novel, mild decellularization strategy which retains the biochemical and biomechanical characteristics of native tendon upon decellularization. We further show that decellularized tendon sections in vitro induce tenogenic differentiation in stem cells and anti-inflammatory response in naïve macrophages. Upon implantation in an Achilles tendon defectin rabbits, we observed that decellularized tendons integrated with the host tissue without signs of tissue rejection or encapsulation. Together, we demonstrate that decellularized tendons produced with our new protocol bear a great potential for tendon tissue regeneration.