Agent for the storage and suspension of cells, especially erythrocytes

• US 5,554,527 A
• Filed: 04/20/1992
• Issued: 09/10/1996
• Est. Priority Date: 11/24/1989
• Status: Expired due to Term

• Current Assignee: Siemens Healthcare Diagnostics Products GmbH (Siemens AG)
• Original Assignee: Behringwerke Aktiengesellschaft (Sanofi )
• Inventors: Fickenscher, Karl
• Primary Examiner(s): Kepplinger, Esther M.
• Assistant Examiner(s): Koh, Choon P.
• Application Number:
  US07/871,575
• Time in Patent Office: 1,604 Days
• Field of Search: 435/240.1, 435/2, 435/7.8, 436/8, 436/18, 424/601, 252/380, 252/397, 514/774, 514/822
• US Class Current: 435/372
• CPC Class Codes: A01N 1/02 Preservation of living parts
• 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: A61K 35/18 Erythrocytes haemoglobin A61K38/42
• CPC Class Codes: G01N 2496/05 containing blood cells or plasma
• CPC Class Codes: G01N 33/68 involving proteins, peptides or amino acids involving lipoproteins G01N33/92
• CPC Class Codes: G01N 33/96 involving blood or serum control standard
• 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 436/108331 Preservative, buffer, anticoagulant or diluent

Abstract

The invention relates to an agent for the preservation, storage and suspension of cells, especially erythrocytes, which contains a chelating agent for multiply charged metal ions, in addition to other substances known to those skilled in the art, such as, for example, electrolytes and sugars.

Claims

1. A method for the preservation and storage of erythrocytes in a suspendable form comprising the steps of:

a) separating erythrocytes from blood containing said erythrocytes; and

b) suspending the separated erythrocytes in an aqueous solution buffered at a pH of 5 to 9, and containing 2 to 50 mmol/l of a chelating agent for the chelation of multiply charged ions, wherein the aqueous solution in step (b) further comprises per liter 3-100 mmol of glucose or fructose, 20-200 mmol of sodium chloride, 10-50 mmol of potassium chloride, 0.5-10 mmol of magnesium chloride, 2-100 mmol of HEPES, 0-100 mmol of sodium phosphate, 10,000-20,000 units of heparin, 1-5 grams of gelatin or polygeline, 2-50 mmol of ATP and 0-1 g of 5-chloro-2-methyl-3[2H]isothiazolone and 2-methyl-3 isothiazolone or chloramphenicol, and wherein the erythrocytes are preserved for at least six months.

2. The method of claim 1 wherein the chelating agent is selected from the group consisting of EDTA, EGTA, and oxalic acid.

3. The method as claimed in claim 1 wherein the pH of the aqueous solution is 6.5 to 7.5.

4. An aqueous solution obtained by the method of claim 1 containing suspendable or suspended erythrocytes.

5. A method for the determination of complement activity which comprises (a) mixing a sample to be tested for complement activity with the aqueous solution of claim 4 and a solution which contains antibodies which specifically bind erythrocytes, and (b) determining lysis time of the erythrocytes as a measure of the complement activity of said sample.

6. A method for the determination of the activity of inhibitors of the complement system which comprises (a) mixing a sample to be tested for the activity of inhibitors with the aqueous solution of claim 4, a solution which contains antibodies which specifically bind erythrocytes, and complement, and (b) determining lysis time of the erythrocytes as a measure of the activity of the inhibitors of said sample.