Sperm suspensions for use in insemination

• US 20050214733A1
• Filed: 03/29/2005
• Published: 09/29/2005
• Est. Priority Date: 03/29/2004
• Status: Active Grant

• Current Assignee: Inguran LLC
• Original Assignee: Monsanto Technology LLC (Bayer AG)
• Inventors: Crowley, Kathleen S., Graham, Jeffrey A., Ludwig, Cindy L.
• Granted Patent: US 7,892,725 B2
• Time in Patent Office: Days
• 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 unsorted spermatozoa, the process comprising

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

b. storing the sperm dispersion.

12. The process of claim 1, wherein the concentration of spermatozoa in the sperm dispersion is at least about 0.04×

10⁶ sperm/ml.

13. The process of claim 1, wherein the concentration of spermatozoa in the sperm dispersion is at least about 1.5×

10⁶ sperm/ml.

14. The process of claim 1, wherein the concentration of spermatozoa in the sperm dispersion is at least about 1×

10⁷ sperm/ml.

15. The process of claim 1, wherein the concentration of spermatozoa in the sperm dispersion is at least about 12×

10⁷ sperm/ml.

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

1, respectively.

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

1.

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

31. The process of claim 1, wherein the dispersion comprises NaHCO₃, KHCO₃, and C₆H₈O₇.H₂O.

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

2. A process for storing sorted spermatozoa, the process comprising

a. forming a sperm dispersion, the sperm dispersion comprising spermatozoa and a composition that induces sperm immotility,

b. sorting the sperm dispersion into separate populations, wherein the spermatozoa of one of the populations comprises at least about 65% X chromosome bearing sperm cells or at least about 65% Y chromosome bearing sperm cells, and

c. storing the one population at a temperature of about −

4°

C. to about 30°

C.

9. The process of claim 2, wherein the sperm dispersion is stored for at least about 24 hours.

10. The process of claim 2, wherein the sperm dispersion is stored for at least about 72 hours.

11. The process of claim 2, wherein the sperm dispersion is stored for at least about 1 week.

16. The process of claim 2, wherein the concentration of spermatozoa in the sperm dispersion is at least about 0.04×

10⁶ sperm/ml.

17. The process of claim 2, wherein the concentration of spermatozoa in the sperm dispersion is at least about 1.5×

10⁶ sperm/ml.

18. The process of claim 2, wherein the concentration of spermatozoa in the sperm dispersion is at least about 1×

10⁷ sperm/ml.

19. The process of claim 2, wherein the concentration of spermatozoa in the sperm dispersion is at least about 12×

10⁷ sperm/ml.

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

1, respectively.

28. The process of claim 2, wherein the composition that induces sperm immotility comprises potassium and optionally sodium, the molar ratio of potassium to sodium being greater than 1.75:

1.

33. The process of claim 2, wherein the sperm dispersion comprises a source of carbonate.

34. The process of claim 2, wherein the dispersion comprises NaHCO₃, KHCO₃, and C₆H₈O₇.H₂O.

35. The process of claim 2, wherein the dispersion comprises a buffer comprising 0.097 moles/L of NaHCO₃, 0.173 moles/L of KHCO₃, 0.090 moles/L C₆H₈O₇.H₂O in water.

39. The process of claim 2, wherein the spermatozoa of one of the populations comprises at least about 70% X chromosome bearing or at least about 70% Y chromosome bearing sperm cells.

40. The process of claim 2, wherein the spermatozoa of one of the populations comprises at least about 80% X chromosome bearing or at least about 80% Y chromosome bearing sperm cells.

41. The process of claim 2, wherein the spermatozoa of one of the populations comprises at least about 90% X chromosome bearing or at least about 90% Y chromosome bearing sperm cells.

42. The process of claim 2, wherein the spermatozoa of one of the populations comprises at least about 95% X chromosome bearing or at least about 95% Y chromosome bearing sperm cells.

3. A process for inseminating a female mammal, the process comprising inseminating a female mammal with a sperm dispersion, the sperm dispersion comprising immotile spermatozoa and a composition that induces sperm immotility.

43. The process of claim 3, wherein the sperm dispersion comprises spermatozoa sorted into separate populations, wherein the spermatozoa of one of the populations comprises at least about 65% X chromosome bearing sperm cells or at least about 65% Y chromosome bearing sperm cells.

44. The process of claim 3, wherein the sperm dispersion comprises spermatozoa sorted into separate populations, wherein the spermatozoa of one of the populations comprises at least about 75% X chromosome bearing sperm cells or at least about 75% Y chromosome bearing sperm cells.

45. The process of claim 3, wherein the sperm dispersion comprises spermatozoa sorted into separate populations, wherein the spermatozoa of one of the populations comprises at least about 85% X chromosome bearing sperm cells or at least about 85% Y chromosome bearing sperm cells.

46. The process of claim 3, wherein the sperm dispersion comprises spermatozoa sorted into separate populations, wherein the spermatozoa of one of the populations comprises at least about 90% X chromosome bearing sperm cells or at least about 90% Y chromosome bearing sperm cells.

47. The process of claim 3, wherein the sperm dispersion comprises spermatozoa sorted into separate populations, wherein the spermatozoa of one of the populations comprises at least about 95% X chromosome bearing sperm cells or at least about 95% Y chromosome bearing sperm cells.

4. A process for providing a fresh sperm dispersion for inseminating a female mammal, the process comprising:

a. forming a sperm dispersion, the sperm dispersion comprising spermatozoa and a composition that induces sperm immotility,

b. placing the sperm dispersion in a container for shipment to a remote location, and

c. shipping the sperm dispersion in the container to a remote location within about 24 hours after forming the sperm dispersion.

6. The process of claim 4, wherein the sperm dispersion is stored at about −

4°

C. to about 30°

C.

7. The process of claim 4, wherein the sperm dispersion is stored at about 0°

C. to about 5°

C.

8. The process of claim 4, wherein the sperm dispersion is stored at about 5°

C. to about 30°

C.

48. The process of claim 4, wherein the sperm dispersion comprises spermatozoa sorted into separate populations, wherein the spermatozoa of one of the populations comprises at least about 65% X chromosome bearing sperm cells or at least about 65% Y chromosome bearing sperm cells.

49. The process of claim 4, wherein the sperm dispersion comprises spermatozoa sorted into separate populations, wherein the spermatozoa of one of the populations comprises at least about 75% X chromosome bearing sperm cells or at least about 75% Y chromosome bearing sperm cells.

50. The process of claim 4, wherein the sperm dispersion comprises spermatozoa sorted into separate populations, wherein the spermatozoa of one of the populations comprises at least about 85% X chromosome bearing sperm cells or at least about 85% Y chromosome bearing sperm cells.

51. The process of claim 4, wherein the sperm dispersion comprises spermatozoa sorted into separate populations, wherein the spermatozoa of one of the populations comprises at least about 90% X chromosome bearing sperm cells or at least about 90% Y chromosome bearing sperm cells.

52. The process of claim 4, wherein the sperm dispersion comprises spermatozoa sorted into separate populations, wherein the spermatozoa of one of the populations comprises at least about 95% X chromosome bearing sperm cells or at least about 95% Y chromosome bearing sperm cells.

53. The process of claim 4, wherein the sperm dispersion is shipped within about 24 hours after the formation of the sperm dispersion.

54. The process of claim 4, wherein the sperm dispersion is shipped within about 48 hours after the formation of the sperm dispersion.

5. A combination comprising:

a. an elongated container for use in the insemination of a female mammal, and

b. a sperm dispersion, the sperm dispersion comprising immotile spermatozoa and a composition that induces sperm immotility, and wherein the sperm dispersion is contained in the elongated container.

20. The combination of claim 5, wherein the concentration of spermatozoa in the sperm dispersion is at least about 0.04×

10⁶ sperm/ml.

21. The combination of claim 5, wherein the concentration of spermatozoa in the sperm dispersion is at least about 1.5×

10⁶ sperm/ml.

22. The combination of claim 5, wherein the concentration of spermatozoa in the sperm dispersion is at least about 1×

10⁷ sperm/ml.

23. The combination of claim 5, wherein the concentration of spermatozoa in the sperm dispersion is at least about 12×

10⁷ sperm/ml.

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

1, respectively.

29. The combination of claim 5, wherein the composition that induces sperm immotility comprises potassium and optionally sodium, the molar ratio of potassium to sodium being greater than 1.75:

1.

36. The combination of claim 5, wherein the sperm dispersion comprises a source of carbonate.

37. The combination of claim 5, wherein the dispersion comprises NaHCO₃, KHCO₃, and C₆H₈O₇.H₂O.

38. The combination of claim 5, wherein the dispersion comprises a buffer comprising 0.097 moles/L of NaHCO₃, 0.173 moles/L of KHCO₃, 0.090 moles/L C₆H₈O₇.H₂O in water.