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Compositions and Methods for Tissue Repair with Extracellular Matrices

Inactive Publication Date: 2011-08-04
RGT UNIV OF CALIFORNIA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0017]In one aspect, the invention provides a therapeutic method for cardiac tissue repair in a subject comprising injecting or implanting a therapeutically effective amount of a composition comprising decellularized extracellular matrix derived from cardiac tissue into a subject in need thereof.
[0018]In another aspect, a composition herein comprises decellularized extracellular matrix derived from sk

Problems solved by technology

Eventually heart failure is onset, and the heart dilates, leading to decreased pumping efficiency.
As there are very few cardiac progenitors in the heart, and these progenitors do not divide readily and regeneration of the heart tissue does not occur naturally.
Current treatments for heart failure rely heavily on invasive surgical procedures and do little to repair damaged heart tissue.
More recently investigated procedures utilize the injection of healthy cells into the left ventricle (LV) infarct wall in an attempt to regenerate the myocardium, although studies have shown poor injected cell survival.
None of these provide a significant amount of the native components of the heart extracellular matrix.

Method used

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  • Compositions and Methods for Tissue Repair with Extracellular Matrices
  • Compositions and Methods for Tissue Repair with Extracellular Matrices
  • Compositions and Methods for Tissue Repair with Extracellular Matrices

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0098]Various studies to treat MI have investigated the injection of cells directly into the infarct wall, although many studies have shown poor survival rates. The objective of this study is to examine the use of a gel as a growth platform for cell adhesion, growth, maturation, and delivery in vivo. It is provided that a gel composed of native heart extracellular matrix tissue can aid in cardiac tissue regeneration by promoting cell survival.

[0099]Female Sprague Dawley rats were enthanized and their hearts decellularized using a procedure modified from Ott et al. (Nature Medicine, 14(2), 213, 2008). Decellularized hearts were then lyophilized, rehydrated, pulverized, and lyophilized again to form a dry powder. The ECM was then minimally digested in pepsin and neutralized, as modified from Freytes et al. (Biomaterials 29: 1630, 2008).

[0100]More specifically, adult female Sprague Dawley rats were heparinized and anesthetized intraperitoneally with pentobarbital. The aorta and pulmona...

example 2

[0109]Cardiomyocytes have been typically cultured on surfaces coated with one, or possibly a few extracellular matrix (ECM) proteins. Yet, in vivo, cardiomyocytes exist in a highly complex extracellular milieu; an ECM that more closely mimics this native environment may be beneficial for cultured cardiomyocyte survival. Here, the use of native cardiac ECM that has been solubilized as a coating for cell culture of neonatal cardiomyocytes is reported.

[0110]Hearts were removed from Sprague-Dawley rats and decellularized using a modified Langendorff setup (modified from Ott et al., 2008). The decellularized hearts were lyophilized, rehydrated, and pulverized after freezing in liquid N2. The ECM was minimally digested in pepsin in 0.01M HCl. After 48 hours, 0.01 M acetic acid was added to make the final concentration of 1 mg / ml.

[0111]Cardiac myocytes were harvested from freshly dissected ventricles of 1 to 2 day old Sprague-Dawley rats using an isolation kit (Cellutron, Highland Park, N....

example 3

[0116]Here, cell coating use has been investigated for native heart extracellular matrix of adult ventricles that have been decellularized and solubilized. The advantages being that native heart ECM may have more components than traditional cell coatings, and be more readily available for use than pretreatment with other cell types.

[0117]Hearts were removed from Sprague-Dawley rats, and decellularized using a modified Langendorff setup (modified from Ott et al, 2008). The decellularized hearts were lyophilized, rehydrated, and pulverized after freezing in liquid nitrogen. The ECM was then digested in pepsin in 0.1M HCl. After 48 hours of digestion, 0.01 M acetic acid was added to dilute to the final concentration of 1 mg / ml.

[0118]Pepsin digestion of the native heart ECM was run in vertical gel electrophoresis in reducing conditions using DTT and compared against laminin (BD Biosciences), and calf skin collagen (Sigma). Gels were stained with Imperial Protein Stain (Pierce). Native h...

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Abstract

Described herein are compositions comprising decellularized cardiac extracellular matrix and therapeutic uses thereof. Methods for treating, repairing or regenerating defective, diseased, damaged or ischemic cells, tissues or organs in a subject, preferably a human, using a decellularized cardiac extracellular matrix of the invention are provided. Methods of preparing cardiomyocyte culture surfaces and culturing cells with absorbed decellularized cardiac extracellular matrix are provided.

Description

CROSS REFERENCES TO RELATED APPLICATIONS[0001]This patent application is a continuation of PCT Application No. PCT / US2009 / 039823, filed Sep. 30, 2009, which claims priority benefit of U.S. Provisional Application No. 61 / 101,332 filed Sep. 30, 2008, each of which is incorporated herein by reference in their entireties.STATEMENT OF GOVERNMENT INTEREST[0002]This invention was made with government support under grant No. OD004309 awarded by National Institutes of Health (NIH). The government has certain rights in the invention.BACKGROUND[0003]Various publications, including patents, published applications, technical articles and scholarly articles are cited throughout the specification. Each of these cited publications is incorporated by reference herein, in its entirety.[0004]Cardiovascular disease is the leading cause of death in the United States. The most common cause of cardiovascular disease is myocardial infarction (MI), which occurs when a coronary artery is occluded. MI results...

Claims

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Application Information

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IPC IPC(8): A61K35/12A61K38/02A61K35/34C12P21/00C12N5/077
CPCA61K35/34A61L27/3633A61L27/367A61L2400/06A61L27/3834A61L2430/20A61P21/00A61P9/00A61P9/06C12N5/0068A61K9/0024A61L27/52
Inventor CHRISTMAN, KARENSINGELYN, JENNIFERDEQUACH, JESSICA
Owner RGT UNIV OF CALIFORNIA
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