Cell preservative solution

• US 5,256,571 A
• Filed: 05/01/1991
• Issued: 10/26/1993
• Est. Priority Date: 05/01/1991
• Status: Expired due to Term

• Current Assignee: Cytyc Corporation (Hologic Incorporated)
• Original Assignee: Cytyc Corporation (Hologic Incorporated)
• Inventors: Oud, Peter S., Lapen, Daniel C., Hurley, Anne A.
• Primary Examiner(s): Robinson, Douglas W.
• Assistant Examiner(s): DADIO, SUSAN
• Application Number:
  US07/694,452
• Time in Patent Office: 909 Days
• Field of Search: 435/1, 435/2, 435/240.1, 435/240.2, 435/800, 436/17, 436/18, 436/826, 436/8
• US Class Current: 436/17
• CPC Class Codes: A01N 1/02 Preservation of living parts
• CPC Class Codes: A01N 1/0221 Freeze-process protecting agents, i.e. substances protecting cells from effects of the physical process, e.g. cryoprotectants, osmolarity regulators like oncotic agents
• CPC Class Codes: A01N 1/0226 Physiologically active agents, i.e. substances affecting physiological processes of cells and tissue to be preserved, e.g. anti-oxidants or nutrients
• CPC Class Codes: C12N 1/04 Preserving or maintaining viable microorganisms immobilised microorganisms C12N11/00
• CPC Class Codes: Y10S 436/826 Additives, e.g. buffers, diluents, preservatives
• CPC Class Codes: Y10T 436/10 Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
• CPC Class Codes: Y10T 436/107497 Preparation composition [e.g., lysing or precipitation, etc.]
• CPC Class Codes: Y10T 436/108331 Preservative, buffer, anticoagulant or diluent

Abstract

An aqueous alcohol buffer solution for substantially ambient, in vitro preservation of mammalian cells for a selected duration. The solution generally contains a water-miscible alcohol in an amount sufficient to fix the sample cells without coagulation, an anti-clumping agent, and a buffer agent to maintain the solution at a pH within a range of four to seven.

Claims

1. An aqueous alcohol-buffer solution for maintaining the structural integrity of mammalian cells in vitro, said solution comprising

A. a water-miscible alcohol in an amount sufficient to fix mammalian cells,

B. an anti-clumping agent in an amount sufficient to prevent the mammalian cells from clumping in said solution, and

C. a buffering agent which maintains said solution, with the mammalian cells, at a pH range of between about two to about seven.

2. The solution of claim 1 wherein said alcohol is selected from the group consisting of ethanol, isopropanol, and methanol.

3. The solution of claim 1 wherein said alcohol is methanol.

4. The solution of claim 1 wherein said anti-clumping agent is a chelating agent selected from the group consisting of ethylenediamine tetraacetic acid and salts thereof.

5. The solution of claim 1 wherein said anti-clumping agent is ethylenediamine tetraacetic acid.

6. The solution of claim 1 wherein said alcohol constitutes about 45 to about 55 percent of said solution.

7. The solution of claim 1 wherein said alcohol constitutes about 50 percent of said solution.

8. The solution of claim 1 wherein said buffering agent is selected from the group consisting of phosphate buffered saline, Tris buffer, sodium acetate, ethylenediamine tetraacetic acid, ethylenediamine tetraacetic acid salts, citric aid and citric acid salts.

9. The solution of claim 1 wherein said buffering agent is sodium acetate.

14. The solution of claim 1 wherein said alcohol constitutes about 20 percent of said solution.

15. The solution of claim 1 wherein said anti-clumping agent is an ethylenediamine tetraacetic acid salt selected from the group consisting of sodium and potassium.

10. An aqueous alcohol-buffer solution for maintaining the structural integrity of mammalian cells in vitro, said solution comprising, by volume,

A. about forty-five to fifty-five percent methanol,

B. about two to four percent acetic acid, and

C. about six to eight percent sodium acetate buffer.

11. The solution of claim 10 comprising about 50 percent methanol.

12. The solution of claim 10 comprising about three percent acetic acid.

13. The solution of claim 10 comprising about seven percent sodium acetate buffer.