Tubular bioengineered smooth muscle structures

Patent metadata

Field	Value
Publication number	US-9675646-B2
Application number	US-201314375818-A
Country	US
Kind code	B2
Filing date	Jan 31, 2013
Priority date	Jan 31, 2012
Publication date	Jun 13, 2017
Grant date	Jun 13, 2017

Abstract

Methods of generating tubular, bioengineered, smooth muscle structures are disclosed as well as bioengineered tissue for tubular organ repair or replacement. The methods can include the steps of obtaining smooth muscle cells; culturing the muscle cells to form a smooth muscle cell construct of directionally oriented smooth muscle cells; disposing the smooth muscle cell construct around a tubular scaffold; and culturing construct and scaffold in a growth media until a smooth muscle cell structure is achieved. The step of obtain smooth muscle cells can further include obtaining autologous smooth muscle cells from a subject. In one preferred embodiment, the muscle cells can first be on a fibrin substrate to form a muscle construct, which is then disposed around a tubular scaffold, for example, a chitosan scaffold. The methods of the present invention can further include connecting two or more tubular structures together to form an elongate composite structure.

First claim

What is claimed is:

1. A method of generating a tubular, bioengineered, gastrointestinal tissue structure comprising:
obtaining smooth muscle cells;
culturing a first population of the smooth muscle cells to form a first annular smooth muscle cell construct of circumferentially oriented s 5 mooth muscle cells;
culturing a second population of the smooth muscle cells to form a second annular smooth muscle cell construct of circumferentially oriented smooth muscle cells;
joining the first and second annular smooth muscle cell constructs together around a tubular scaffold, wherein the tubular scaffold comprises chitosan; and
culturing the constructs and scaffold in a growth media until a gastrointestinal tissue structure is achieved that exhibits a travelling wave of contractions through the first and second annular smooth muscle cell constructs in response to a contractile stimulus.

2. The method of claim 1 , wherein the step of obtaining smooth muscle cells comprises obtaining smooth muscle cells from a subject.

3. The method of claim 1 , wherein the step of culturing the first and second population of smooth muscle cells comprises culturing at least one of said first or second population of the smooth muscle cells in a collagen suspension.

4. The method of claim 1 , wherein the first and second annular smooth muscle cell constructs are joined together by a surgical glue.

5. A tubular, gastrointestinal tissue structure formed by the method of claim 1 .

6. The method of claim 1 wherein the first and second annular smooth muscle cell constructs are joined together with a gelatin glue.

7. The method of claim 1 wherein the first and second annular smooth muscle cell constructs are joined together with a fibrin glue.

8. The method of claim 1 wherein the first and second annular smooth muscle cell constructs are also bonded to the tubular scaffold.

9. The method of claim 1 wherein the step of culturing the constructs and scaffold in a growth media further comprises culturing the constructs and scaffold in a bioreactor.

10. The method of claim 1 wherein the step of culturing the constructs and scaffold in a growth media further comprises culturing the constructs and scaffold in a low-shear, rotating wall bioreactor.

11. A gastrointestinal tissue structure comprising:
a first population of smooth muscle cells forming a first annular smooth muscle cell structure;
a second population of smooth muscle cells forming a second annular smooth muscle cell structure; and
a tubular scaffold comprising chitosan;
wherein the first and second annular smooth muscle cell structures are disposed about the scaffold and joined together such that the resulting gastrointestinal smooth muscle tissue structure exhibits circumferentially oriented smooth muscle cells, basal tone and cholinergic contractions in response to a contractile stimulus.

12. The tissue structure of claim 11 wherein the structure also exhibits a travelling wave of contractions through the first and second annular smooth muscle cell constructs in response to a contractile stimulus.

Assignees

• Univ Wake Forest Health Sciences

Inventors

• Bitar Khalil

Classifications

• C12N2533/72

  Chitin, chitosan · CPC title

• C12N2533/54

  Collagen; Gelatin · CPC title

• C12N5/0661

  Smooth muscle cells · CPC title

• C12N5/0068

  General culture methods using substrates (for specific animal cell type C12N5/06) · CPC title

• A61L2430/30

  for muscle reconstruction · CPC title