Compositions and methods for improving the quality of processed sperm
View Patent ↗The present invention relates to compositions and methods for the handling of processed sperm including samples that are freshly collected, those transported as fresh samples, samples that are frozen and thawed, those sorted into one or more subpopulations, and those that are otherwise processed or handled that impose trauma on the cell. Trauma can reduce the motility, fertility, viability and overall integrity of the sperm and reduce the ability to fertilize, produce an embryo and a healthy offspring. The present invention relates to novel compounds that can be added to the sperm cell sample to reduce the traumatic effects of physical stress during mild as well as extensive sperm cell processing, methods of using the compounds in standard sperm processing procedures, the end products made from these methods including sperm and embryos, as well as methods of using those end products in assisted reproductive biology techniques in animals.
1. A method of fertilizing one or more eggs, comprising the steps of:
providing a sperm cell sample in a media making a sperm cell composition, wherein the media comprises vitamin B12, or a vitamin B12 vitamer, at a concentration of about 0.01 mg/ml to 1 mg/ml and alpha-ketoglutarate; and
contacting the one or more eggs with one or more sperm cells from the sperm cell composition.
2. The method of claim 1 , wherein the concentration of the vitamin B12, or the vitamin B12 vitamer, in the media is selected from the group consisting of: 0.01 to 0.25 mg/ml; 0.01 to 0.5 mg/ml; 0.05 to 0.1 mg/ml; 0.05 to 1.0 mg/ml; 0.1 to 0.25 mg/ml; 0.1 to 0.5 mg/ml; 0.1 to 1 mg/ml; 0.15 to 0.45 mg/ml; 0.15 to 0.5 mg/ml; 0.25 to 0.35 mg/ml; 0.25 to 0.5 mg/ml; 0.25 to 1 mg/ml; 0.35 to 0.5 mg/ml; 0.35 to 1 mg/ml; and 0.5 to 1 mg/ml.
3. The method of claim 1 , wherein the concentration of the vitamin B12, or the vitamin B12 vitamer, in the media is selected from the group consisting of: about 0.05 mg/ml; about 0.1 mg/ml; about 0.15 mg/ml; about 0.25 mg/ml; about 0.35 mg/ml; about 0.45 mg/ml; and about 0.5 mg/ml.
4. The method of claim 1 , wherein the concentration of the vitamin B12, or the vitamin B12 vitamer, in the media is about 0.15 mg/ml, about 0.25 mg/ml or about 0.35 mg/ml.
5. The method of claim 1 , wherein the concentration of the vitamin B12, or the vitamin B12 vitamer, in the media is about 0.25 mg/ml.
6. The method of claim 1 , wherein the concentration of the alpha-ketoglutarate in the media is about 0.15 mg/ml to 0.5 mg/ml.
7. The method of claim 1 , wherein the sperm cell sample has been sex-sorted.
8. The method of claim 1 , wherein the sperm cell sample has been processed by ablating unwanted cells.
9. The method of claim 1 , further comprising the step of freezing and thawing the sperm cell sample or the sperm cell composition.
10. The method of claim 1 , wherein the sperm cell sample is derived from a human, bovine, swine, ovine, equine, deer, elk, buffalo, canine, feline, chimpanzee or gorilla, or a whale, dolphin or other marine mammal.
11. A method of fertilizing one or more eggs, comprising the steps of:
providing a sex-sorted sperm cell sample, or a sperm cell sample processed by ablating unwanted cells, in a media making a sperm cell composition, wherein the media comprises vitamin B12, or a vitamin B12 vitamer, at a concentration of about 0.01 mg/ml to 1 mg/ml; and
contacting the one or more eggs with one or more sperm cells from the sperm cell composition.
12. The method of claim 11 , wherein the concentration of the vitamin B12, or the vitamin B12 vitamer, in the media is about 0.15 mg/ml, about 0.25 mg/ml or about 0.35 mg/ml.
13. The method of claim 11 , wherein the concentration of the vitamin B12, or the vitamin B12 vitamer, in the media is about 0.25 mg/ml.
14. The method of claim 11 , wherein the media further comprises alpha-ketoglutarate.
15. The method of claim 11 , further comprising the step of freezing and thawing the sperm cell sample or the sperm cell composition.
16. The method of claim 11 , wherein the sperm cell sample is derived from a human, bovine, swine, ovine, equine, deer, elk, buffalo, canine, feline, chimpanzee or gorilla, or a whale, dolphin or other marine mammal.