Tissue-engineered bowel constructs

Patent metadata

Field	Value
Publication number	US-10828143-B2
Application number	US-201715808338-A
Country	US
Kind code	B2
Filing date	Nov 9, 2017
Priority date	Dec 30, 2015
Publication date	Nov 10, 2020
Grant date	Nov 10, 2020

Abstract

Methods are disclosed for forming tissue engineered, tubular bowel constructs from intestinal circular smooth muscle cells and enteric neural progenitor cells. The intestinal smooth muscle cells and neural progenitor cells can be seeded on a mold with a surface texture that induces longitudinal alignment of the intestinal smooth muscle cells and co-cultured until an innervated aligned smooth muscle sheet is obtained. The innervated smooth muscle sheet can then be wrapped around a tubular scaffold to form an intestinal tissue construct.

First claim

The invention claimed is:

1. A method of forming a tissue engineered bowel construct comprising:
isolating intestinal circular smooth muscle cells from an intestinal donor source,
isolating enteric neural progenitor cells from an intestinal donor source,
seeding the isolated intestinal circular smooth muscle cells on a plate with grooves that induces longitudinal alignment of the intestinal circular smooth muscle cells,
adding the isolated enteric neural progenitor cells to the intestinal circular smooth muscle cells on the plate,
co-culturing the intestinal circular smooth muscle cells and the enteric neural progenitor cells until an innervated aligned smooth muscle sheet is obtained,
wrapping the innervated aligned smooth muscle sheet around a hollow tubular scaffold to form an innervated bowel construct.

2. The method of claim 1 , wherein the plate is comprised of a non-adherent surface with a wavy microtopography.

3. The method of claim 2 wherein the surface comprises a silicon elastomer.

4. The method of claim 2 wherein the surface is coated with a biological agent.

5. The method of claim 2 wherein the biological agent coating comprises laminin.

6. The method of claim 1 , wherein the enteric neural progenitor cells are suspended in a collagen/laminin gel.

7. The method of claim 1 , wherein steps of isolating the intestinal circular smooth muscle cells and enteric neural progenitor cells further comprise obtaining each type of cell from a single subject.

8. The method of claim 1 , wherein the co-culturing step comprises culturing cells in a collagen suspension.

9. The method of claim 1 , wherein the tubular scaffold comprises a biodegrabable material.

10. The method of claim 1 , wherein the tubular scaffold comprises chitosan.

11. The method of claim 1 , further comprising implanting the bowel construct into an intestinal system of a subject.

12. The method of claim 1 , wherein the method further comprises connecting two or more innervated aligned smooth muscle sheets together to form a composite structure.

13. A tubular bowel construct comprising:
a hollow tubular scaffold with a central lumen;
an innervated intestinal tissue sheet of longitudinally aligned smooth muscle cells wrapped circumferentially around the tubular scaffold, and formed by seeding isolated intestinal circular smooth muscle cells on a plate with grooves that induces longitudinal alignment of the intestinal circular smooth muscle cells, adding the isolated enteric neural progenitor cells to the intestinal circular smooth muscle cells on the plate, and co-culturing the intestinal circular smooth muscle cells and the enteric neural progenitor cells until an innervated aligned smooth muscle sheet is obtained,
the construct exhibiting directionally oriented smooth muscle cells, basal tone and choleric contractions in response to a contractile stimulus.

14. The construct of claim 13 , wherein the hollow tubular scaffold comprises a biodegradable material.

15. The construct of claim 13 , wherein the hollow tubular scaffold comprises chitosan.

16. The construct of claim 13 , wherein the tubular bowel construct is 1 cm to 100 cm long.

17. The construct of claim 13 , wherein the tubular bowel construct is 5 cm to 50 cm long.

18. The construct of claim 13 , wherein the tubular bowel construct has an internal diameter from about 0.1 cm to 50 cm.

19. The construct of claim 13 , wherein the tubular bowel construct has an internal diameter from 0.5 cm to 20 cm.

Assignees

• Univ Wake Forest Health Sciences

Inventors

• Bitar Khalil

Classifications

• C12N2502/1347

  Smooth muscle cells · CPC title

• C12N2501/11

  Epidermal growth factor [EGF] · CPC title

• C12N2502/088

  neural stem cells · CPC title

• C12N5/0697

  Artificial constructs associating cells of different lineages, e.g. tissue equivalents (blood vessels C12N5/0691) · CPC title

• C12N2501/115

  Basic fibroblast growth factor (bFGF, FGF-2) · CPC title