Lyophilization of cells

• US 5,045,446 A
• Filed: 07/11/1989
• Issued: 09/03/1991
• Est. Priority Date: 08/26/1988
• Status: Expired due to Fees

• Current Assignee: COBE Laboratories Incorporated (Terumo Corporation)
• Original Assignee: Cryopharm Corp.
• Inventors: Goodrich, Raymond P. Jr., Williams, Christine M.
• Primary Examiner(s): Rosen, Sam
• Application Number:
  US07/378,349
• Time in Patent Office: 784 Days
• Field of Search: 435/2, 435/240.1, 424/533
• US Class Current: 435/2
• CPC Class Codes: A01N 1/02 Preservation of living parts
• CPC Class Codes: A01N 1/0205 Chemical aspects
• 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

Abstract

A process and medium are disclosed for the lyophilization of cells which comprises the use of solutions including monosaccharide hexoses and pentoses, and/or biocompatible amphipathic polymers to permit the reconstitution of viable cells.

Claims

1. A process for the lyophilization of cells having a cell membrane, comprising immersing a plurality of cells in a water solution which includes a monosaccharide, which is capable of permeating the membrane of the cells, in a concentration of from about 7.0 to 37.5%;

freezing the solution; and

drying the frozen cells by sublimation of the water.

2. The process of claim 1 wherein the monosaccharide is selected from the group consisting of pentoses and hexoses.

3. The process of claim 1 or 2 wherein the monosaccharide is selected from the group consisting of xylose, glucose, ribose, mannose and fructose.

4. The process of claim 3 wherein the monosaccharide is present in the solution in a concentration of about 23%.

5. A process for the lyophilization of cells, comprising:

immersing a plurality of cells in a buffered solution which includes;

a monosaccharide which is present in the solution in a concentration of from about 7.0 to 37.5%, and

a polymer having a molecular weight of from about 1K to about 360K which is present in a concentration of from about 0.7% up to saturation in the solution;

freezing the solution; and

drying the cells by sublimation of the water.

6. The process of claim 5 wherein said polymer is amphipathic.

7. The process of claim 5 wherein said polymer has a molecular weight in the range of about 5K to about 80K.

8. The process of claim 5, 6 or 7 wherein the monosaccharide is selected from the group consisting of pentoses and hexoses.

9. The process of claim 8 wherein the monosaccharide is selected from the group consisting of xylose, glucose, ribose, mannose and fructose.

10. The process of claim 8 wherein the polymer is selected from the group consisting of polyvinylpyrrolidone and dextran.

11. The process of claim 10 wherein the polymer is polyvinylpyrrolidone.

12. A process for the lyophilization of cells, comprising:

immersing a plurality of cells in a buffered solution which includes;

a monosaccharide which is present in the solution in a concentration of from about 7.0 to 37.5%, and

a polymer having a molecular weight of from about 1K to about 360K which is present in a concentration of from about 0.7% up to saturation in the solution;

freezing the solution; and

drying the cells by sublimation of the water.

13. The process of claim 12 wherein said polymer is amphipathic.

14. The process of claim 12 wherein said polymer has a molecular weight in the range of about 5K to about 80K.

15. The process of claim 12, 13 or 14 wherein the monosaccharide is selected from the group consisting of pentoses and hexoses.

16. The process of claim 15 wherein the monosaccharide is selected from the group consisting of xylose, glucose, ribose, mannose and fructose.

17. The process of claim 15 wherein the polymer is selected from the group consisting of polyvinylpyrrolidone and dextran.

18. The process of claim 17 wherein the polymer is polyvinylpyrrolidone.