Methods and compositions to improve germ cell and embryo survival and function

• US 6,140,121 A
• Filed: 10/17/1996
• Issued: 10/31/2000
• Est. Priority Date: 10/19/1995
• Status: Expired due to Term

• Current Assignee: Bio-Origyn LLC
• Original Assignee: Advanced Reproduction Technologies Inc.
• Inventors: Oliver, Sylvia Adams, Ellington, Joanna E.
• Primary Examiner(s): Stucker, Jeffrey
• Assistant Examiner(s): NELSON, BRETT L
• Application Number:
  US08/733,227
• Time in Patent Office: 1,475 Days
• Field of Search: 435/374, 435/366, 435/404
• US Class Current: 435/374
• CPC Class Codes: A01N 1/021 Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
• 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: A61K 35/52 Sperm; Prostate; Seminal fluid; Leydig cells of testes
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: C12N 2500/60 Buffer, e.g. pH regulation, osmotic pressure
• CPC Class Codes: C12N 5/061 Sperm cells, spermatogonia

Abstract

Sperm, oocyte, and embryo survival and function is improved in vivo or in vitro by the use of a polysaccharide containing arabinose, galactose and/or hexuronic acid. In particular, a nonspermicidal lubricant containing such a polysaccharide (e.g., gum arabic, pectin, or galacturonic acid) increases the fertilization potential of the sperm during coitus, artificial insemination or sperm collection. Similarly, a freezing medium containing a polysaccharide containing arabinose, galactose and/or hexuronic acid enhances sperm, oocyte, or embryo viability.

Claims

1. A method for isolation of sperm with improved function, comprising:

(a) contacting a sample containing sperm with a solution comprising a hexuronic acid monomer or with a solution comprising a polymer wherein said polymer comprises hexuronic acid and arabinose or galactose or both arabinose and galactose, to form a mixture; and

(b) subjecting the mixture in step (a) to a condition sufficient to separate sperm from the sample, thereby isolating sperm with improved function.

2. The method of claim 1 wherein the solution of step (a) comprises said polymer.

4. The method of claim 2 wherein the solution comprises said polymer.

3. A method for washing sperm to obtain sperm with improved function, comprising contacting a sample containing sperm with a solution comprising a hexuronic acid monomer or with a solution comprising a polymer wherein said polymer comprises hexuronic acid and arabinose or galactose or both arabinose and galactose, and removing the solution, thereby obtaining, sperm with improved function.

5. A method for extending sperm to obtain sperm with improved function, comprising contracting a sample containing sperm with a solution comprising a hexuronic acid monomer or with a solution comprising a polymer wherein said polymer comprises hexuronic acid and arabinose or galactose or both arabinose and galactose, thereby obtaining sperm with improved function.

6. The method of claim 5 wherein the solution comprises said polymer.

7. A sperm wash medium comprising a solution comprising a balanced salt solution and a hexuronic acid monomer or a solution comprising a balanced salt solution and a polymer wherein said polymer comprises a hexuronic acid and arabinose or galactose or both arabinose and galactose, at a concentration sufficient to improve sperm function.

8. The medium of claim 7 wherein the solution comprises a balanced salt solution and said polymer.

9. A sperm isolation medium for isolation of sperm by centrifugation, comprising a solution comprising a density gradient material and a hexuronic acid monomer or a solution comprising a density gradient material and a polymer wherein said polymer comprises hexuronic acid and arabinose or galactose or both arabinose and galactose.

10. The medium of claim 9 wherein the solution comprises a density gradient material and said polymer.

11. A sperm isolation medium for the isolation of sperm by swim-up method, comprising a solution comprising a macromolecule and a hexuronic acid monomer or a solution comprising a macromolecule and a polymer wherein said polymer comprises hexuronic acid and arabinose or galactose or both arabinose and galactose.

12. The medium of claim 11 wherein the solution comprises a macromolecule and said polymer.

13. An extending or culturing medium comprising a solution comprising a balanced salt solution and a hexuronic acid monomer or a solution comprising a balanced salt solution and a polymer wherein said polymer comprises hexuronic acid and arabinose or galactose or both arabinose and galactose, at a concentration sufficient to improve sperm, oocyte, embryo, or embryonic stem cell (ESC) function.

14. The medium of claim 13 wherein the solution comprises a balanced salt solution and said polymer.

15. A method for reducing loss of functional sperm during storage of sperm in a refrigerated, frozen or vitrified state, comprising the steps of:

(a) combining a hexuronic acid monomer, or combining a polymer wherein said polymer comprises hexuronic acid and arabinose or galactose or both arabinose and galactose, with a sample containing sperm, wherein said hexuronic acid monomer or said polymer is in an amount effective to reduce said loss; and

(b) storing said sample in a refrigerated, frozen or vitrified state.

16. The method of claim 15 wherein in step (a) said polymer is combined with a sample containing sperm.