Engineered tissues for in vitro research uses, arrays thereof, and methods of making the same

Patent metadata

Field	Value
Publication number	US-12372515-B2
Application number	US-202017013165-A
Country	US
Kind code	B2
Filing date	Sep 4, 2020
Priority date	Sep 12, 2011
Publication date	Jul 29, 2025
Grant date	Jul 29, 2025

Abstract

Disclosed are living, three-dimensional tissue constructs for in vitro scientific and medical research, arrays thereof, and methods of making said tissues and arrays.

First claim

What is claimed is:

1. An in vitro method of producing a living, three-dimensional tissue construct comprising:
a. bioprinting at least one semi-solid or solid bio-ink comprising endothelial cells and a different cell type in the same or separate bio-inks; and
b. incubating the at least one semi-solid or solid bio-ink in vitro to cohere and form the living, three-dimensional tissue construct comprising a microvascular network consisting of endothelial cells that is located within the construct and is surrounded by cells that are not part of the microvascular network;
wherein the tissue construct does not comprise an artery or vein.

2. The method of claim 1 , wherein the bio-ink comprises multicellular aggregates comprising mammalian cells.

3. The method of claim 1 , wherein the bioprinting further comprises arranging a plurality of cell types relative to each other to create a planar geometry in at least one layer.

4. The method of claim 1 , comprising bioprinting at least one layer that is compositionally or architecturally distinct from at least one other layer to create a laminar geometry.

5. The method of claim 1 , wherein the tissue construct is at least about 25 μm in its smallest dimension at the time of bioprinting, wherein the tissue construct is no greater than about 3 cm in its largest dimension at the time of bioprinting, or both.

6. The method of claim 1 , wherein the endothelial cells, the different cell type, or both are differentiated cells.

7. The method of claim 1 , wherein the endothelial cells, the different cell type, or both are non-differentiated cells.

8. The method of claim 1 , comprising isolating the different cell type from a tissue selected from the group consisting of: liver, gastrointestinal, pancreatic, kidney, lung, tracheal, vascular, skeletal muscle, cardiac, skin, smooth muscle, connective tissue, corneal, genitourinary, breast, reproductive, endothelial, epithelial, fibroblast, neural, adipose, bone, bone marrow, cartilage, mesothelial, endocrine, stromal, lymph, blood, endoderm, ectoderm, mesoderm, or a combination thereof.

9. The method of claim 1 , comprising isolating the endothelial cells from a tissue selected from the group consisting of: blood, blood vessel, lymphatic vessel, tissue of the digestive tract, tissue of the genitourinary tract, adipose tissue, tissue of the respiratory tract, tissue of the reproductive system, mesoderm-derived tissue, bone marrow, and umbilical tissue.

10. The method of claim 1 , wherein the tissue construct is a vascular wall segment.

11. The method of claim 1 , comprising producing multiple tissue constructs and constructing an array from the multiple tissue constructs.

12. The method of claim 1 , wherein the at least one semi-solid or solid bio-ink is cylindrical.

13. The method of claim 12 , wherein the bioprinting comprises continuous deposition of the at least one semi-solid or solid bio-ink.

14. The method of claim 13 , wherein the bioprinting comprises extruding the at least one semi-solid or solid bio-ink through a dispense tip of a bioprinter.

15. The method of claim 14 , wherein the dispense tip is connected to a reservoir comprising the at least one semi-solid or solid bio-ink.

16. The method of claim 1 , wherein the at least one semi-solid or solid bio-ink is not bioprinted into a pre-formed scaffold.

17. The method of claim 1 , comprising bioprinting multiple layers of the at least one semi-solid or solid bio-ink.

18. The method of claim 17 , wherein each layer is in direct contact with at least one other layer.

Assignees

• Organovo Inc

Inventors

• Murphy Keith
• Khatiwala Chirag
• Dorfman Scott
• Shepherd Benjamin
• Presnell Sharon
• Robbins Justin

Classifications

• G01N33/5082

  Supracellular entities, e.g. tissue, organisms · CPC title

• C12N2513/00

  3D culture · CPC title

• C12N2502/28

  Vascular endothelial cells · CPC title

• C12N2502/27

  Lung cells, respiratory tract cells · CPC title

• C12N5/0697

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