IP Library Granted Patent US 8,123,924
Granted Patent B2
US 8,123,924 · App. 10/574,911 · Granted Feb 28, 2012

Sperm cell separation by electrophoresis

Assignee: Newcastle Innovation Limited
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Quick Facts
Patent No.
US 8,123,924
App. No.
10/574,911
Granted
Feb 28, 2012
Kind
B2
Abstract

A process for separating a sperm type from a sperm population by electrophoresis comprising subjecting the sperm population to an electric potential such that a sperm type is separated from a sperm population through an ion-permeable barrier.

Claims (30)

1. A process for separating a sperm type from a sperm population in a sperm sample by electrophoresis comprising subjecting the sperm population to an electric potential such that a sperm type move through an ion-permeable barrier and the sperm type is separated from the sperm population through the ion-permeable barrier.

2. The process according to claim 1 wherein the sperm type has a desired characteristic selected from the group consisting of motility, robustness, gender, fertilizing potential, and combinations thereof.

3. The process according to claim 2 wherein the sperm type has fertilizing potential.

4. The process according to claim 1 wherein the sperm type has an undesired characteristic of poor motility.

5. The process according to claim 1 comprising:

providing a sample containing sperm to a sample chamber of electrophoresis apparatus comprising a first electrolyte chamber; a second electrolyte chamber; a first sample chamber disposed between the first electrolyte chamber and the second electrolyte chamber; a second sample chamber disposed adjacent to the first sample chamber; a first ion-permeable barrier disposed between the first sample chamber and the second sample chamber; a second ion-permeable barrier disposed between the first electrolyte chamber and the first sample chamber; a third ion-permeable barrier disposed between the second sample chamber and the second electrolyte chamber; and electrodes disposed in the first and second electrolyte chambers; and applying an electric potential between the electrodes causing at least one sperm type in the first or second sample chamber to move through the first ion-permeable barrier into the other of the first or second sample chamber.

6. The process according to claim 5 wherein one or more of the first, second, or third ion-permeable barriers are electrophoresis membranes having a characteristic average pore size and pore size distribution.

7. The process according to claim 6 wherein all of the ion-permeable barriers are electrophoresis membranes having a characteristic average pore size and pore size distribution.

8. The process according to claim 5 wherein the first ion-permeable barrier is a large pore sized membrane.

9. The process according to claim 8 wherein the first ion-permeable barrier is a polycarbonate membrane having a pore size of about 1 to about 10 μm.

10. The process according to claim 1 wherein the sperm sample contains at least two populations of sperm.

11. The process according to claim 1 wherein the electric potential is applied until at least one sperm type reaches a desired purity.

12. The process according to claim 1 wherein electrophoresis is carried out with a voltage range from 1 to 200 V.

13. The process according to claim 1 wherein electrophoresis is carried out with an applied current of up to about 75 mA.

14. The process according to claim 1 wherein electrophoresis is carried out with a field strength of 1 to 100 V/cm.

15. The process according to claim 14 wherein the field strength is from 16 to 20 V/cm.

16. The process according to claim 1 wherein electrophoresis is carried out from several seconds to about 15 minutes in an apparatus having a sample volume of about 100μ to about 4 ml.

17. The process according to claim 1 wherein sperm is diluted in buffer having a concentration of between 1 to 100 mM.

18. The process according to anyone of claims 1 to 17 wherein the sperm sample has a sperm concentration of between about 0.1 and 250×10 6 /ml.

19. The process according to claim 18 wherein the sperm concentration is between 15×10 6 /ml and 140×10 6 /ml are processed.

20. The process according to claim 1 wherein at least about 50% of the sperm type remains viable or substantially unchanged after separation.

21. The process according to claim 20 wherein at least about 60% of the sperm type remains viable or substantially unchanged after separation.

22. The process according to claim 21 wherein at least about 70% of the sperm type remains viable or substantially unchanged after separation.

23. The process according to claim 22 wherein at least about 80% of the sperm type remains viable or substantially unchanged after separation.

24. The process according to claim 23 wherein at least about 90% of the sperm type remains viable or substantially unchanged after separation.

25. A method of fertilizing an ovum comprising contacting the ovum with a sperm type separated by the process of claim 1 .

26. The process of claim 1 , wherein 3.5 hours after treatment the separated sperm type has a motility value which is 94±10% of the motility value of untreated sperm.

27. The process according to claim 1 wherein at least about 50% of the sperm type remains substantially unchanged after separation.

28. The process according to claim 1 further comprising selecting the sperm type wherein the selected sperm type has a desired characteristic selected from the group consisting of morphological normality, low levels of DNA damage, low levels of reactive oxygen species generation, and combinations thereof.

29. The process of claim 1 further comprising selecting the sperm type wherein the selected sperm type has an undesired characteristic selected from the group consisting of poor morphology, high levels of DNA damage and high levels of reactive oxygen species generation.

Assignments (2)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jan 6, 2012
From: THE UNIVERSITY OF NEWCASTLE
To: NEWCASTLE INNOVATION LIMITED
Reel/Frame 027494/0648 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 29, 2011
From: GRADIPORE LIMITED
To: UNIVERSITY OF NEWCASTLE
Reel/Frame 026200/0579 →
Priority Claims (1)
AU 2003905474 · Oct 7, 2003 · national
Continuity (1)
Related Publication 20090101507A1 · Apr 23, 2009