Decellularized tissue engineered constructs and tissues

• US 6,962,814 B2
• Filed: 08/16/2001
• Issued: 11/08/2005
• Est. Priority Date: 08/16/2000
• Status: Expired due to Term

• Current Assignee: Duke University
• Original Assignee: Duke University
• Inventors: Prabhakar, Vikas, Niklason, Laura, Mitchell, Shannon, Koh, Jennifer
• Primary Examiner(s): Guzo, David
• Assistant Examiner(s): Sullivan, Daniel M.
• Application Number:
  US09/931,506
• Publication Number:
  US 20020115208A1
• Time in Patent Office: 1,545 Days
• Field of Search: 514/2, 514/44, 435/70.1, 435/402, 435/395, 424/93.1, 424/93.2, 424/93.21, 623/23.76
• US Class Current: 435/402
• 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 27/3839 characterised by the site of application in the body materials for artificial blood vessels A61L27/507; materials for use in artificial skin A61L27/60
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: C12N 2533/30 Synthetic polymers
• CPC Class Codes: C12N 2533/40 Polyhydroxyacids, e.g. polymers of glycolic or lactic acid (PGA, PLA, PLGA); Bioresorbable polymers
• CPC Class Codes: C12N 2533/90 Substrates of biological origin, e.g. extracellular matrix, decellularised tissue
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06

Abstract

New methods for producing tissue engineered constructs and engineered native tissues are disclosed. The methods include producing a tissue engineered construct by growing cells in vitro on a substrate and then decellularizing the construct to produce a decellularized construct consisting largely of extracellular matrix components. The construct can be used immediately or stored until needed. The decellularized construct can be used for further tissue engineering, which may include seeding the construct with cells obtained from the intended recipient of the construct. During any of the growth phases required for production of the construct, the developing construct may be subjected to various tissue engineering steps such as application of mechanical stimuli including pulsatile forces. The methods also include producing an engineered native tissue by harvesting tissue from an animal or human, performing one or more tissue engineering steps on the tissue, and subjecting the tissue to decellularization. The decellularized, engineered native tissue may then be subjected to further tissue engineering steps.

Claims

1. A construct for use as a tissue engineering scaffold or for implanting into a subject comprising:

a tissue engineered construct comprising a 3-dimensional proteinaceous extracellular matrix that has been subjected to decellularization, wherein the tissue engineered construct was formed by seeding a substrate with cells and maintaining said cells under conditions suitable for growth of the cells, whereby a proteinaceous extracellular matrix surrounding said cells is formed, wherein the substrate comprises a collagen sponge.

2. A construct for use as a tissue engineering scaffold or for implanting into a subject comprising:

a tissue engineered construct comprising a 3-dimensional proteinaceous extracellular matrix that has been subjected to decellularization, wherein the tissue engineered construct was formed by seeding a substrate with cells and maintaining said cells under conditions suitable for growth of the cells, whereby a proteinaceous extracellular matrix surrounding said cells is formed, wherein the construct is treated so as to remove substantially all of the substrate.

3. A construct for use as a tissue engineering scaffold or for implanting into a subject comprising:

a tissue engineered construct comprising a 3-dimensional proteinaceous extracellular matrix that has been subjected to decellularization, wherein the tissue engineered construct was formed by seeding a substrate with cells and maintaining said cells under conditions suitable for growth of the cells, whereby a proteinaceous extracellular matrix surrounding said cells is formed, wherein the cells comprise porcine cells.

4. A construct for use as a tissue engineering scaffold or for implanting into a subject comprising:

a tissue engineered construct comprising a 3-dimensional proteinaceous extracellular matrix that has been subjected to decellularization, wherein the tissue engineered construct was formed by seeding a substrate with cells and maintaining said cells under conditions suitable for growth of the cells, whereby a proteinaceous extracellular matrix surrounding said cells is formed, wherein the cells comprise tumor cells.