Decellularized mammalian extracellular matrix morsels, methods making and methods of using same

Patent metadata

Field	Value
Publication number	US-2023348859-A1
Application number	US-202318339980-A
Country	US
Kind code	A1
Filing date	Jun 22, 2023
Priority date	Dec 23, 2020
Publication date	Nov 2, 2023
Grant date	—

Abstract

Described are decellularized human extracellular matrix (ECM) morsels for use in tissue regeneration and repair.

First claim

We claim:

1 . A customized flowable composition comprising decellularized human heart extracellular matrix (ECM), wherein said customized flowable composition comprises ECM comprising a desired physical or biochemical/chemical profile differing from the physical or biochemical/chemical profile of in-vivo derived adult human heart ECM in collagen and sulphated glycosaminoglycans (sGAG) content.

2 . A method of producing customized decellularized human heart extracellular matrix (ECM) morsels, the method comprising:
seeding cultured human heart cells into micro-wells, wherein the cultured human heart cells generate an ECM microtissue comprised of the cultured cells and ECM produced from the cultured cells; collecting the ECM microtissue as spheroid particles; and decellularizing the ECM microtissue; wherein said customized decellularized ECM morsels comprise a desired physical or biochemical/chemical profile differing from the physical or biochemical/chemical profile of in-vivo derived adult human heart ECM in collagen and sulphated glycosaminoglycans (sGAG) content.

3 . The method of claim 2 , wherein said decellularizing comprises mixing the microtissue with at least one of a detergent, a buffer, a DNAse, or an RNase.

4 . The method of claim 2 , wherein said ECM microtissue comprises spherical particles having a diameter of less than or equal to 800 μm.

5 . The method of claim 2 , wherein said cells comprise recombinant human cells.

6 . The method of claim 2 , wherein said cells comprise at least one of cardiac fibroblasts, cardiac myocytes and cardiac microvascular endothelial cells.

7 . A customized flowable composition comprising decellularized human heart extracellular matrix (ECM) made by the method comprising:
seeding cultured human heart cells into micro-wells, wherein the cultured cells generate an ECM microtissue comprised of the cultured cells and an ECM; collecting the ECM microtissue as spheroid particles; and decellularizing the ECM microtissue; wherein said customized flowable composition comprises ECM comprising a desired physical or biochemical/chemical profile differing from the physical or biochemical/chemical profile of in-vivo derived adult human heart ECM in collagen and sulphated glycosaminoglycans (sGAG) content.

8 . The composition of claim 7 , wherein said decellularizing comprises mixing the cells with at least one of a detergent, a buffer, a DNAse, or an RNase.

9 . The composition of claim 8 , wherein said method further comprises passing the mixture through a syringe.

10 . The composition of claim 9 , wherein said syringe comprises a 27 G needle.

11 . The composition of claim 7 , wherein said cells comprise recombinant human cells.

12 . The composition of claim 7 , wherein said cells comprise at least one of cardiac fibroblasts, cardiac myocytes and cardiac microvascular endothelial cells.

13 . The method of claim 2 , wherein said micro-wells are scaffold-free.

14 . The composition of claim 7 , wherein said micro-wells are scaffold-free.

15 . The method of claim 2 , wherein said modifying comprises adjusting at least one of cell culture media composition, culturing time, oxygen level, or the presence or amount of additional biological factors.

16 . The method of claim 2 , wherein said additional biological factors comprise at least one of a growth factor, a cytokine, and a drug.

17 . A method of producing customized decellularized human heart extracellular matrix (ECM) morsels, the method comprising:
seeding cultured human heart cells into micro-wells, wherein the cultured cells generate an ECM microtissue comprised of the cultured cells and an ECM, wherein said ECM microtissue comprises spherical particles having a diameter of less than or equal to 800 μm; collecting the ECM microtissue; and decellularizing the ECM microtissue; wherein said customized decellularized human heart ECM morsels comprise a desired physical or biochemical/chemical profile differing from the physical or biochemical/chemical profile of in-vivo derived adult human heart ECM in collagen and sulphated glycosaminoglycans (sGAG) content.

18 . The customized flowable composition of claim 1 , wherein the mechanical stiffness of the ECM is comparable to healthy human heart tissue.

19 . The method of claim 2 , wherein the mechanical stiffness of the ECM is comparable to healthy human heart tissue.

20 . The customized flowable composition of claim 7 , wherein the mechanical stiffness of the ECM is comparable to healthy human heart tissue.

21 . The method of claim 17 , wherein the mechanical stiffness of the ECM is comparable to healthy human heart tissue.

Assignees

• Univ Brown

Inventors

• Ip Blanche C
• Morgan Jeffrey R

Classifications

• A61L27/50

  Materials characterised by their function or physical properties {, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials} · CPC title

• A61L27/3683

  subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment · CPC title

• C12N5/0657 Primary

  Cardiomyocytes; Heart cells · CPC title

• C12N2533/90

  Substrates of biological origin, e.g. extracellular matrix, decellularised tissue · CPC title

• C12N2501/998

  Proteins not provided for elsewhere · CPC title