Tissue extracellular matrix particles and applications

Patent metadata

Field	Value
Publication number	US-2016237390-A1
Application number	US-201415024139-A
Country	US
Kind code	A1
Filing date	Sep 24, 2014
Priority date	Sep 24, 2013
Publication date	Aug 18, 2016
Grant date	—

Abstract

An apparatus in the form of a chip is provided wherein the apparatus is prepared with decellularized extracellular matrix from various tissues and can be used to investigate the cellular interactions between the ECM and the various cell types. Three dimensional culture methods for investigating decellularized extracellular matrix from various tissues and interactions with various mammalian cell types are also provided. Methods of use of cells grown using the apparatus and methods disclosed are also provided.

First claim

1 . An apparatus for culturing cells comprising:
a first functionalized substrate having at least one functionalized surface; a gel pad disposed on the functionalized surface of the first functionalized substrate; an array comprising a plurality of discrete layers of collagen positioned on top of the gel pad, each having a defined area and being substantially aligned with one another and defining a space between one another; and a layer of decellularized extracellular matrix particles (DECM) positioned on top of at least one or more of the discrete layers of collagen, wherein the DECM is capable of supporting cellular growth.

2 . The apparatus of claim 1 wherein the functionalized substrate is glass.

3 . The apparatus of claim 1 , wherein the gel pad comprises a polymerized biocompatible gel.

4 . The apparatus of claim 1 , wherein the discrete layers of collagen comprise collagen I.

5 . The apparatus of claim 1 , wherein the discrete layers of collagen are circular in shape.

6 . The apparatus of claim 5 , wherein the discrete layers of collagen have a diameter of between 1 to 10 mm.

7 . The apparatus of any of claim 1 , wherein the DECM particles are derived from one or more different tissues.

8 . The apparatus of claim 7 , wherein the tissues are selected from the group consisting of kidney, lung, liver, spleen, bladder, skeletal muscle, cartilage, bone, heart, intestine, tendon, and brain.

9 . The apparatus of claim 1 , wherein the DECM particles are derived from one or more different species of mammal.

10 . A process for forming spheroid aggregates of mammalian stem cells and decellularized extracellular matrix (DECM) particles comprising:
a) preparing a suspension of DECM particles in a suitable growth media; b) preparing a suspension of mammalian stem cells in the same suitable growth media; c) preparing a mixture of a) and b) at a ratio in a range of 10:1 to 1:10 v/v DECM particle solution:mammalian cells; d) suspending the mixture of c) in a hanging drop culture for a period of between 2 days and 7 days; e) replacing the growth media with a suitable induction media; and f) allowing the culture to grow for period of time sufficient to produce a spheroid aggregate comprising mammalian stem cells and DECM.

11 . The process of claim 10 , further comprising the addition of one or more growth factors or cytokines to the media of e).

12 . The process of claim 10 , wherein the DECM particles are derived from one or more different tissues.

13 . The process of claim 12 , wherein the tissues are selected from the group consisting of kidney, lung, liver, spleen, bladder, skeletal muscle, cartilage, bone, heart, intestine, tendon, and brain.

14 . The process of claim 10 , wherein the DECM particles are derived from one or more different species of mammal.

15 . The process of claim 10 , wherein the mammalian stem cells are selected from the group consisting of adipose stem cells, mesenchymal stem cells, cardiac stem cells, hepatic stem cells, retinal stem cells, and epidermal stem cells.

16 . A spheroid aggregate of mammalian stem cells and DECM particles made using the process of claim 10 .

17 . A method for identifying the interaction of mammalian stem cells with differing types of extracellular matrix in vitro comprising:
a) preparing an apparatus of claim 1 , or using the process of claim 10 , with DECM particles from one or more different tissues; b) obtaining a sample of mammalian stem cells of interest; c) placing a sufficient amount of the cells of interest of a) in the apparatus claim 1 with suitable growth media, or using the process of claim 10 ; d) culturing the mammalian stem cells of interest for a sufficient period of time; and f) comparing the effect of the DECM particles from one or more different tissues on the growth of the mammalian stem cells of interest.

18 . A method of implanting spheroid aggregates of mammalian stem cells and decellularized extracellular matrix (DECM) particles in a subject comprising:
a) identifying a subject in need of spheroid aggregates of mammalian stem cells and decellularized extracellular matrix (DECM) particles; b) identifying a site in the subject in need of implantation of spheroid aggregates of mammalian stem cells and decellularized extracellular matrix (DECM) particles; and c) implanting the spheroid aggregates of mammalian stem cells and decellularized extracellular matrix (DECM) particles in the subject at the identified site.

19 . The method of claim 18 , wherein the spheroid aggregates of mammalian stem cells and decellularized extracellular matrix (DECM) particles are made using the process of claim 10 .

Assignees

• Univ Johns Hopkins

Inventors

• Beachley Vincent
• Gibson Matt
• Elisseeff Jennifer

Classifications

• C12M25/14 Primary

  Scaffolds; Matrices (in general C12N5/0068) · CPC title

• A61K35/545

  Embryonic stem cells; Pluripotent stem cells; Induced pluripotent stem cells; Uncharacterised stem cells · CPC title

• C12M25/01

  Drops · CPC title

• C12M23/20

  Material Coatings (immunocoatings C12M25/00) · CPC title

• C12M23/04

  Flat or tray type, drawers (C12M23/10, C12M23/12, C12M23/16 take precedence) · CPC title