Organ/tissue decellularization, framework maintenance and recellularization

A method of regenerating an organ from an acellular organ scaffold for a subject in need comprising:

• (a) providing an acellular organ scaffold, wherein the organ is an ex vivo organ that is diseased, injured, damaged, or discarded for a medical reason, and wherein the organ is decellularized by a method comprising the steps of:
• (i) perfusing the ex vivo organ with at least 50 volumes of a detergent, wherein the volume is the volume of the organ being perfused;
• and
• (ii) rinsing the ex vivo organ with at least 50 volumes of a neutral buffer, wherein the perfusing and rinsing are performed using at least two different fluid inlets and at least one different fluid outlet, thereby resulting in a decellularized organ;
• (b) contacting the acellular organ scaffold of step (a) with a cell characteristic of the acellular organ scaffold;
• (c) incubating the acellular organ scaffold and cell ex vivo under ramping perfusion conditions to produce an incubated organ scaffold, wherein the perfusion comprises delivery of at least 1×108 cells at a rate spanning 2 ml/minute to 20 ml/minute;
• and
• (d) implanting the incubated organ scaffold into the subject and establishing at least two vascular connections of the scaffold vasculature and the subject vasculature, thereby producing a regenerated organ from an acellular organ scaffold.