Granted Patent B2

US 12,553,089 · App. 17/613,825 · Granted Feb 17, 2026

Inventors: Sanford D. Markowitz (Pepper Pike, OH); Helen Moinova (Beachwood, OH); David Wurtman (New York, NY)

Assignee: Case Western Reserve University

C12Q1/6886C12N15/1003C12Q1/6806C12Q2600/154

Abstract

Claims (23)

1 . A method of preserving methylation patterns of a methylated DNA sequence in human esophageal cells, comprising:

treating the human esophageal cells with an aqueous storage solution at room temperature;

storing the human esophageal cells in the aqueous storage solution for at least 24 hours, and

extracting DNA from the human esophageal cells after the storing of the human esophageal cells in the aqueous storage solution,

wherein the aqueous storage solution comprises from 40% to less than 100% methanol, and wherein the methanol is peroxide free or at a level less than or equal to 0.001%,

wherein the aqueous storage solution is free of metal ions,

wherein the DNA from the human esophageal cells comprises a methylated DNA sequence,

wherein the methylated DNA sequence comprises methylation patterns, and

wherein at least 60% of the methylation patterns present in the methylated DNA sequence in the human esophageal cells prior to the storing in the aqueous storage solution are preserved when stored for at least 24 hours in the aqueous storage solution.

2 . The method of claim 1 , comprising storing the human esophageal cells in the aqueous storage solution for at least two weeks, wherein the methylation patterns of the DNA of the human esophageal cells are preserved by at least 60% in comparison to the methylation patterns of the DNA from the human esophageal cells prior to storage in the aqueous storage solution.

3 . The method of claim 1 , comprising storing the human esophageal cells in the aqueous storage solution for at least 24 hours at room temperature, at 4° C. or at temperatures ranging between −30° C. to 50° C., wherein the methylation patterns of the DNA of the human esophageal cells in the aqueous storage solution are preserved by at least 65% in comparison to the methylation patterns in the human esophageal cells prior to storage in the aqueous storage solution.

4 . The method of claim 1 , wherein the aqueous storage solution consists essentially of an admixture of from 40% methanol to less than 100% methanol and water.

5 . The method of claim 1 , comprising storing the human esophageal cells in the aqueous storage solution at room temperature for at least 24 hours to 2 years or at 4° C. for at least 24 hours to 2 years, wherein the methylation patterns of the DNA of the human esophageal cells in the aqueous storage solution are preserved by at least 60% in comparison to the methylation patterns of the DNA from the human esophageal cells prior to storage in the aqueous storage solution.

6 . The method of claim 1 , comprising storing the human esophageal cells in the aqueous storage solution at temperatures ranging between −30° C. to 50° C. for from at least 24 hours to 2 years, wherein the methylation patterns of the DNA of the human esophageal cells in the aqueous storage solution are preserved by at least 60% in comparison to the methylation patterns of the DNA from the human esophageal cells prior to storage in the aqueous storage solution.

7 . A method of preserving methylation patterns of a methylated DNA sequence, comprising:

treating human esophageal cells with an aqueous storage solution at room temperature;

storing the human esophageal cells in the aqueous storage solution for at least 24 hours at ambient temperature, and

extracting DNA from the human esophageal cells after the storing of the human esophageal cells in the aqueous storage solution,

wherein the aqueous storage solution comprises from 40% to less than 100% methanol,

wherein the aqueous storage solution is free of metal ions,

wherein the DNA from the human esophageal cells comprises a methylated DNA sequence,

wherein the methylated DNA sequence comprises methylation patterns, and

wherein at least 60% of the methylation patterns present in the methylated DNA sequence in the human esophageal cells prior to the storing in the aqueous storage solution are preserved when stored for at least 24 hours in the aqueous storage solution at ambient temperature ranging between −30° C. to 50° C.

Assignments (4)

Continuity (2)

References Cited (125)