Decellularization and recellularization of whole organs

Patent metadata

Field	Value
Publication number	US-10918765-B2
Application number	US-201414783923-A
Country	US
Kind code	B2
Filing date	Apr 10, 2014
Priority date	Apr 11, 2013
Publication date	Feb 16, 2021
Grant date	Feb 16, 2021

Abstract

The present invention provides systems and methods for the decellularization and recellularization of tissue segments. In certain instances the invention comprises coating or sealing decellularized tissue segments with a cross-linked alginate hydrogel. The present invention also provides a decellularization kit, which may be used to generate decellularized tissue segments for high throughput studies. Also included are compositions and methods of tissue sealants comprising methacrylated alginate.

First claim

What is claimed:

1. A method of recellularizing a decellularized lung tissue, comprising the steps of:
decellularizing an isolated lung tissue sample to form a decellularized lung tissue, wherein the isolated lung tissue sample is a whole organ or a portion thereof;
applying a first coating to at least a portion of the decellularized lung tissue, wherein the first coating comprises a hydrogel sealing agent comprising a hydrogel solution comprising alginate;
applying a second coating to the portion of tissue, wherein the second coating comprises one or more crosslinking agents that crosslink the alginate coated on the decellularized lung tissue; and
contacting the decellularized lung tissue with at least one cell type;
wherein the hydrogel sealing agent coating prevents leakage of liquids and gases from the decellularized isolated tissue sample.

2. The method of claim 1 , wherein the alginate is crosslinked by exposing the hydrogel sealing agent to at least one crosslinking agent selected from:
one or more compounds selected from the group consisting of eosin Y, triethanolamine, calcium chloride; ammonium persulfate (APS) and tetramethylethylenediamine (TEMED), glutaraldehyde, an epoxide, oxidized dextran, p-azidobenzoyl hydrazide, N-[α.-maleimidoacetoxy]succinimide ester, p-azidophenyl glyoxal monohydrate, bis-[β-(4-azidosalicylamido)ethyl]disulfide, bis[sulfosuccinimidyl]suberate, dithiobis[succinimidyl proprionate, disuccinimidyl suberate, 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS), riboflavin and any combination thereof; and
one or more energy sources selected from the group consisting of visible light irradiation, blue light irradiation, ultraviolet irradiation, and any combination thereof.

3. The method of claim 1 , wherein at least one property of the hydrogel sealing agent is controlled by at least one selected from the group consisting of a degree of alginate crosslinking, a degree of alginate methacrylation, and a degree of methacrylated alginate crosslinking.

4. The method of claim 1 , wherein the alginate is methacrylated at one or more methacrylation sites.

Assignees

• Univ Of Vermont And State Agricultural College

Inventors

• Wagner Darcy
• Weiss Daniel J
• Bonenfant Nicholas
• Oldinski Rachael
• Fenn Spencer

Classifications

• C09D105/04

  Alginic acid; Derivatives thereof · CPC title

• C08B37/0084

  Guluromannuronans, e.g. alginic acid, i.e. D-mannuronic acid and D-guluronic acid units linked with alternating alpha- and beta-1,4-glycosidic bonds; Derivatives thereof, e.g. alginates · CPC title

• A61F17/00

  First-aid kits · CPC title

• A61L27/38

  containing added animal cells (organs or tissue containing native cells A61L27/36) · CPC title

• A61L24/08

  Polysaccharides {(A61L24/043 takes precedence)} · CPC title