Engineered intestinal tissue and uses thereof

Patent metadata

Field	Value
Publication number	US-12545897-B2
Application number	US-202217695546-A
Country	US
Kind code	B2
Filing date	Mar 15, 2022
Priority date	Nov 10, 2016
Publication date	Feb 10, 2026
Grant date	Feb 10, 2026

Abstract

Disclosed are methods of assessing the ability of a candidate therapeutic agent to reverse, reduce or prevent intestinal injury by a potential toxic agent using a three-dimensional, engineered, bioprinted, biological intestinal tissue model. Also disclosed are methods of assessing the effect of an agent on intestinal function, the method comprising contacting the agent with a three-dimensional, engineered, bioprinted, biological intestinal tissue model.

First claim

What is claimed is:

1 . A method of making an intestinal tissue model, the method comprising:
a. depositing a layer comprising adult primary intestinal myofibroblasts at a density of at least about 5×10 6 cells per milliliter onto a biocompatible surface; and b. depositing a layer comprising adult primary intestinal epithelial cells onto and in contact with the apical surface of the layer comprising adult primary intestinal myofibroblasts to produce the intestinal tissue model.

2 . The method of claim 1 , wherein at least one of the layer comprising adult primary intestinal myofibroblasts and the layer comprising adult primary intestinal epithelial cells is deposited by bioprinting, ink-jet printing, extrusion, microvalve printing, or microsolenoid valve printing.

3 . The method of claim 1 , wherein the adult primary intestinal myofibroblasts and/or the adult primary intestinal epithelial cells are deposited as part of a bio-ink.

4 . The method of claim 3 , wherein the bio-ink comprises a hydrogel.

5 . The method of claim 4 , wherein the hydrogel is collagen.

6 . The method of claim 1 , further comprising depositing at least one type of immune cell.

7 . The method of claim 6 , wherein the at least one type of immune cell is a myeloid cell and/or lymphoid cell.

8 . The method of claim 7 , wherein the myeloid cell and/or lymphoid cell is a monocyte, macrophage, neutrophil, basophil, eosinophil, dendritic cell, megakaryocyte, or a combination thereof.

9 . The method of claim 6 , wherein the at least one type of immune cell is deposited: as part of at least one of the layer comprising adult primary intestinal myofibroblasts and the layer comprising adult primary intestinal epithelial cells, on top of the layer comprising adult primary intestinal epithelial cells, or below the layer comprising adult primary intestinal myofibroblasts.

10 . The method of claim 6 , wherein the at least one type of immune cell is deposited as a layer or compartment within the intestinal tissue model.

11 . The method of claim 1 , further comprising inducing a phenotype of an intestinal disorder or injury in the intestinal tissue model by: contacting the intestinal tissue model with a treatment, compound, or infectious agent that gives rise to the phenotype.

12 . The method of claim 11 , wherein the phenotype comprises fibrosis and fibrotic scar formation.

13 . The method of claim 11 , wherein the intestinal tissue model is a model of Crohn's disease, ulcerative colitis, or inflammatory bowel disease.

14 . The method of claim 1 , wherein at least one of the layer comprising adult primary intestinal myofibroblasts and the layer comprising adult primary intestinal epithelial cells comprises cells from a donor with a disease phenotype.

15 . The method of claim 14 , wherein the disease phenotype is celiac disease, Crohn's disease, ulcerative colitis, irritable bowel syndrome, hemorrhoids, diverticulitis, inflammatory bowel disease, microscopic colitis, lymophocytic colitis, collagenous colitis, endocrine disorders, metabolic disorders, obesity, diabetes, dyslipidemia, intestinal cancer, or colorectal cancer.

16 . The method of claim 1 , wherein at least one of the layer comprising adult primary intestinal myofibroblasts and the layer comprising adult primary intestinal epithelial cells is substantially a monolayer.

17 . The method of claim 1 , wherein a biocompatible membrane is in contact with the basolateral surface of the layer comprising adult primary intestinal myofibroblasts.

18 . The method of claim 1 , wherein the intestinal tissue model is at least two cell layers thick.

19 . The method of claim 1 , comprising depositing the layers in at least one first region and at least one second region, wherein the at least one first region comprises a layer comprising adult primary intestinal myofibroblasts from a healthy donor and a layer comprising adult primary intestinal epithelial cells from a healthy donor, and the at least one second region comprises cells from a diseased donor in at least one of the layer comprising adult primary intestinal myofibroblasts and the layer comprising adult primary intestinal epithelial cells.

20 . The method of claim 1 , further comprising culturing the intestinal tissue model for at least 3 days in cell culture media.

21 . The method of claim 1 , wherein the intestinal tissue model comprises a layered architecture, polarized epithelial morphology, and physiological barrier function.

22 . The method of claim 21 , wherein the layered architecture, polarized epithelial morphology, and physiological barrier function are capable of being-maintained for over two weeks in culture.

23 . The method of claim 1 , wherein the intestinal tissue model:
(i) exhibits at least one of the following:
a. apical staining of villin;
b. tight junctions;
c. an apical brush border;
d. villi-like structures on the epithelial surface;
e. a basal lamina between the layer of interstitial tissue and layer of epithelial cells;
f. secretion of mucus;
g. expression of CYP3A4;
h. expression of p-glycoprotein;
i. expression of glucagon-like peptide-I;
j. expression of BCRP;
k. enteroendocrine cells; or
l. goblet cells;
(ii) does not comprise a fully mature, perfusable vasculature; (iii) does not comprise red blood cells; (iv) is not innervated by the central nervous system; or (v) a combination thereof.

24 . The method of claim 1 , wherein the intestinal tissue model comprises a tumor, tumor fragment, tumor cell, or immortalized cell.

Assignees

• Organovo Inc

Inventors

• Retting Kelsey Nicole
• Nguyen Deborah Lynn Greene
• Madden Lauran

Classifications

• C12N5/0697 Primary

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

• C12N5/0634

  Cells from the blood or the immune system · CPC title

• A61K35/38

  Stomach; Intestine; Goblet cells; Oral mucosa; Saliva · CPC title

• G01N33/5088

  of vertebrates · CPC title

• C12N2535/00

  Supports or coatings for cell culture characterised by topography · CPC title