Organ/tissue decellularization, framework maintenance and recellularization
Methods for decellularizing organs and tissues in vitro and in vivo are provided, as are methods of maintaining organ and tissue frameworks and methods of recellularizing organs and tissues, thereby providing an approach to needed organs or tissues.
1. A method of decellularizing a human organ that is diseased, injured, damaged, or discarded for a medical reason comprising:
(a) Perfusing an ex vivo organ that is diseased, injured, damaged, or discarded for a medical reason with at least 50 volumes of a detergent, wherein the volume is the volume of the organ being perfused; and
(b) 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.
2. The method according to claim 1 wherein the organ is selected from the group consisting of kidney, pancreas, and liver.
3. The method according to claim 1 wherein the flow rate of the two fluid inlets is about equal.
4. The method according to claim 1 further comprising an initial step of delivering distilled water to the organ at a rate of about 12 ml per minute for at least 10 hours.
5. The method according to claim 1 further comprising delivering at least one liter of 0.0025 w/w % DNase solution to the detergent-treated organ prior to rinsing the organ.
6. The method according to claim 1 wherein the detergent is sodium dodecyl sulfate or 214-(2,4,4-trimethylpentan-2-yl)phenoxylethanol.
7. The method according to claim 1 further comprising exposing the organ to a bactericidal agent.
8. The method according to claim 7 wherein the bactericidal agent is povidone-iodine.
9. The method according to claim 1 wherein the neutral buffer contains 10 U/ml heparin.
10. The method according to claim 1 wherein at least one fluid inlet and the fluid outlet are derived from different vessel systems.
11. The method according to claim 10 wherein the at least one fluid inlet is an artery and the fluid outlet is a vein or duct.
12. The method according to claim 1 wherein the organ is diseased.
13. The method according to claim 1 wherein the organ is damaged.
14. The method according to claim 1 wherein the organ has been discarded for a medical reason.
15. The method according to claim 1 further comprising attaching the organ scaffold to an antibody product specifically binding to the cell or to an anti-thrombotic composition.
16. The method according to claim 15 wherein the organ scaffold is attached to an antibody product.
17. The method according to claim 16 wherein the antibody product comprises an antigen binding site for CD31.