Methods and compositions for organ decellularization

• US 20030215945A1
• Filed: 06/18/2003
• Published: 11/20/2003
• Est. Priority Date: 12/29/1999
• Status: Abandoned Application

• Current Assignee: Children's Medical Center Corporation
• Original Assignee: Children's Medical Center Corporation
• Inventors: Atala, Anthony
• Application Number:
  US10/464,165
• Publication Number:
  US 20030215945A1
• Time in Patent Office: Days
• US Class Current: 435/376
• CPC Class Codes: A61L 2430/26 for kidney reconstruction
• CPC Class Codes: A61L 2430/40 Preparation and treatment of biological tissue for implantation, e.g. decellularisation, cross-linking
• CPC Class Codes: A61L 27/3604 characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
• CPC Class Codes: A61L 27/3687 characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
• CPC Class Codes: A61L 27/3691 characterised by physical conditions of the treatment, e.g. applying a compressive force to the composition, pressure cycles, ultrasonic/sonication or microwave treatment, lyophilisation
• CPC Class Codes: A61L 27/38 containing added animal cells organs or tissue containing native cells A61L27/36

Abstract

The invention is directed to methods for producing a decellularized organ or part of an organ. A decellularized organ is produced using an isolated organ mechanically agitated to remove cellular membranes surrounding the isolated organ without destroying the interstitial structure of the organ. After the cellular membrane is removed, the isolated organ is exposed to a solubilizing fluid that extracts cellular material without dissolving the interstitial structure of the organ. A washing fluid is used to remove the solubilized components, leaving behind a decellularized organ.

Claims

1. A solubilizing fluid for tissue decellularization comprising:

an alkaline solution selected from the group consisting of sulphates, acetates, carbonates, bicarbonates, hydroxides, ammonium salts and combinations thereof; and

a non-ionic detergent wherein the concentration of detergent is in the range of about 0.006% (w/v) to 2.0% (w/v).

2. The composition of matter of claim 1, wherein the non-ionic detergent is selected from the group consisting of Triton X-100, Triton N-101, Triton X-114, Triton X-405, Triton X-705, and Triton DF-16, monolaurate (Tween 20), monopalmitate (Tween 40), monooleate (Tween 80), polyoxethylene-23-lauryl ether (Brij 35), polyoxyethylene ether W-1 (Polyox), sodium cholate, deoxycholates, CHAPS, saponin, n-Decyl β

-D-glucopuranoside, n-heptyl β

-D glucopyranoside, n-Octyl α

-D-glucopyranoside, Nonidet P-40, and combinations thereof.

3. The composition of matter of claim 1, wherein the alkaline solution comprises ammonium hydroxide and the detergent comprises Triton X-100.