	US 12,337,078 B2
	Organ/tissue decellularization, framework maintenance and recellularization
Giuseppe Orlando, Winston-Salem, NC (US); and Shay Soker, Greensboro, NC (US)
Assigned to WAKE FOREST UNIVERSITY HEALTH SCIENCES, Winston-Salem, NC (US)
Filed by WAKE FOREST UNIVERSITY HEALTH SCIENCES, Winston-Salem, NC (US)
Filed on Feb. 17, 2023, as Appl. No. 18/170,669.
Application 18/170,669 is a division of application No. 14/610,902, filed on Jan. 30, 2015, granted, now 11,648,335.
Claims priority of provisional application 61/934,429, filed on Jan. 31, 2014.
Prior Publication US 2023/0310708 A1, Oct. 5, 2023
Int. Cl. A61L 27/36 (2006.01); A61L 27/38 (2006.01); A61L 27/54 (2006.01)

CPC A61L 27/3633 (2013.01) [A61L 27/3604 (2013.01); A61L 27/3687 (2013.01); A61L 27/3808 (2013.01); A61L 27/3834 (2013.01); A61L 27/54 (2013.01); A61L 2300/256 (2013.01); A61L 2300/42 (2013.01); A61L 2430/26 (2013.01); A61L 2430/28 (2013.01); A61L 2430/40 (2013.01)]

9 Claims

1. 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 1×10⁸cells 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.