Process for storing a sperm dispersion

• US 7,892,725 B2
• Filed: 03/29/2005
• Issued: 02/22/2011
• Est. Priority Date: 03/29/2004
• Status: Active Grant

• Current Assignee: Inguran LLC
• Original Assignee: Inguran LLC
• Inventors: Crowley, Kathleen S., Graham, Jeffrey A., Ludwig, Cindy L.
• Primary Examiner(s): Gitomer; Ralph
• Assistant Examiner(s): GOUGH, TIFFANY MAUREEN
• Application Number:
  US11/092,338
• Publication Number:
  US 20050214733A1
• Time in Patent Office: 2,156 Days
• Field of Search: 435/2
• US Class Current: 435/2
• 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 1/0226 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 35/52 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 2500/05 Inorganic components
• CPC Class Codes: C12N 5/061 Sperm cells, spermatogonia
• CPC Class Codes: C12N 5/0611 Primordial germ cells, e.g. embryonic germ cells [EG]
• CPC Class Codes: G01N 1/30 Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis

Abstract

Processes of storing sorted and unsorted spermatozoa, in the form of a sperm dispersion, having reduced motility relative to endogenous ejaculated sperm are disclosed. The immotile spermatozoa of the dispersion tend to have a greater capacity for enduring the rigors associated with storage, transportation, and fertilization procedures. Processes for providing such sperm dispersions, for inseminating a female mammal with such sperm dispersions, as well as compositions and combinations comprising the sperm dispersions, are also disclosed.

Claims

1. A process for storing gender enriched spermatozoa, the process comprising the steps of:

a. forming an immotile sperm dispersion, the immotile sperm dispersion comprising spermatozoa, a composition that induces sperm immotility, and an antibiotic;

b. sorting the immotile sperm dispersion, while rendered immotile by the composition that induces sperm immotility, into a gender enriched dispersion of X chromosome bearing sperm or Y chromosome bearing sperm; and

c. storing the gender enriched sperm dispersion.

2. The process of claim 1 wherein the composition that induces sperm immotility comprises potassium and sodium , the molar ratio of potassium and sodium being greater than 1:

1, respectively.

3. The process of claim 1 wherein the composition that induces sperm immotility comprises potassium and sodium, the molar ratio of potassium and sodium being greater than 1.75:

1.

4. The process of claim 1 wherein the composition that induces sperm immotility comprises potassium and sodium, the molar ration of potassium and sodium being greater than 2:

1.

5. The process of claim 1 wherein the composition that induces sperm immotility comprises potassium and sodium, the molar ratio of potassium and sodium being greater than 3:

1.

6. The process of claim 1 wherein the sperm dispersion comprises a source of carbonate.

7. The process of claim 1 wherein the immotile sperm dispersion comprises NaHCO₃, KHCO₃, and C₆H₈O₇.H₂O in water.

8. The process of claim 1 wherein the immotile dispersion comprises a buffer comprising 0.097 moles/L of NaHCO₃, 0.173 moles/L of KHCO₃, and 0.090 C₆H₈O₇.H₂O in water.

9. The process of claim 1 wherein the immotile sperm dispersion further comprises a DNA selective dye.

10. The process of claim 9 wherein the DNA selective dye is selected from the group consisting of Hoechst 3342, Hoechst 3358, and SYBR-14.

11. The process of claim 1 wherein immotile dispersion is stored in an enlongate container for use in the insemination of a female mammal.

12. The process of claim 11 wherein the elongate container for use in the insemination of a female mammal has a volume from about 0.1 cc to about 100 cc.

13. The process of claim 1 wherein the immotile sperm dispersion further comprises one selected from the group of:

vitamin K, Lipoic Acid, and Pyruvate, and combinations thereof.

14. The process of claim 1 wherein prior to the step of storing a cryoextender is added to the gender enriched sperm dispersion.

15. The process of claim 14 wherein the cryoextender further comprises a motility inhibitor.

16. The process of claim 1 wherein the gender enriched sperm dispersion is stored at a temperature between 5°

C. and 30°

C.

17. The process of claim 1 wherein the gender enriched sperm dispersion is stored at a temperature between −

196°

C. and 30°

C.

18. The process of claim 1 further comprising the step of fertilizing an embryo with the gender enriched sperm dispersion.

19. The process of claim 18 wherein the fertilization occurs within 24 hours of the formation of the gender enriched sperm dispersion.

20. The process of claim 19 wherein the fertilization occurs within 96 hours of the formation of the gender enriched sperm dispersion.