Methods of preserving mesenchymal stem cells

Patent metadata

Field	Value
Publication number	US-11701389-B2
Application number	US-201716083661-A
Country	US
Kind code	B2
Filing date	Mar 9, 2017
Priority date	Mar 9, 2016
Publication date	Jul 18, 2023
Grant date	Jul 18, 2023

Abstract

This disclosure relates to methods of preserving mesenchymal stromal/stem cells (MSCs) for use in clinical applications. In certain embodiments, this disclosure relates to methods of preserving MSCs comprising mixing MSCs with interferon-gamma prior to cryopreserving, freezing, or cooling the MSCs to a temperature below zero degrees Celsius.

First claim

The invention claimed is:

1. A method of cryopreserving mesenchymal stem cells comprising
mixing mesenchymal stem cells with interferon gamma at a concentration of 20 ng/mL in culture medium for 48 hours providing mixed mesenchymal stem cells, and cooling the mixed mesenchymal stem cells to a temperature below zero degrees Celsius providing frozen interferon gamma exposed mesenchymal stem cells, wherein mixing is prior to cooling the mixed mesenchymal stem cells, and
thawing the frozen interferon gamma exposed mesenchymal stem cells to a temperature above zero degrees Celsius providing thawed interferon gamma exposed mesenchymal stem cells that express indoleamine 2,3-dioxygenase (IDO) mRNA with greater than a 100-fold change in concentration when compared to cryopreserved mesenchymal stem cells that were not mixed with interferon gamma prior to freezing.

2. The method of claim 1 , wherein the temperature is less than negative 50 degrees Celsius.

3. The method of claim 1 , wherein the mesenchymal stem cells are in a cryoprotectant during cooling.

4. The method of claim 3 wherein the cryoprotectant comprises a component selected from monosaccharide, a disaccharide, trehalose, glycerol, ethylene glycol, and propylene glycol, or combinations thereof.

5. The method of claim 3 wherein the cryoprotectant comprises a polyampholyte.

6. The method of claim 1 further comprising mixing the thawed interferon gamma exposed mesenchymal stem cells with a component selected from interferon-gamma, interleukin-1beta, interleukin-1alpha, transforming growth factor-beta, interleukin-6, interleukin-10, prostaglandin-E2, hepatocyte growth factor, tumor necrosis factor-alpha, or combinations thereof.

7. The method of claim 1 further comprising mixing the thawed interferon gamma exposed mesenchymal stem cells with interferon-gamma optionally in combination with a cytokine selected from interleukin-1beta, interleukin-1 alpha, and tumor necrosis factor-alpha.

8. The method of claim 1 further comprising purifying, plating, passaging, and harvesting the thawed mesenchymal stem cells providing regenerated mesenchymal stem cells.

Assignees

• Univ Emory
• Childrens Healthcare Atlanta Inc

Inventors

• Galipeau Jacques
• Chinnadurai Raghavan

Classifications

• A01N1/162

  Temperature processes, e.g. following predefined temperature changes over time · CPC title

• A01N1/126

  Physiologically active agents, e.g. antioxidants or nutrients · CPC title

• A01N1/125

  Freeze protecting agents, e.g. cryoprotectants or osmolarity regulators · CPC title

• A01N1/124

  Disinfecting agents, e.g. antimicrobials · CPC title

• A61K35/28 Primary

  Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells · CPC title