Compositions and methods for improving the quality of processed sperm
The present invention generally relates to compositions and methods for the handling of processed sperm populations including samples that are freshly collected, those transported as fresh samples, as well as 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 sperm cell. Such trauma can reduce the motility, fertility, viability and overall integrity of the sperm and reduce the sperm's ability to fertilize an egg, grow into a health embryo and produce 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 processing sperm cells comprising the steps of:
a) forming a sperm cell composition comprising the sperm cells and alpha-ketoglutarate, wherein the alpha-ketoglutarate is at a concentration ranging from 0.01 mg/ml to 5 mg/ml in the sperm cell composition;
b) analyzing the sperm cells with a flow cytometer; and
c) sex-sorting the sperm cells or ablating unwanted sperm cells among the sperm cells in the sperm cell composition with a laser.
2. The method of claim 1 , wherein the step of sorting comprises sorting the sperm cell composition to form at least one gender enriched population of X-chromosome bearing or Y-chromosome bearing sperm cells.
3. The method of claim 1 , wherein the alpha-ketoglutarate is at a concentration ranging from 0.01 mg/ml to 1 mg/ml in the sperm cell composition.
4. The method of claim 3 , wherein the alpha-ketoglutarate is at a concentration ranging from 0.15 mg/ml to 0.5 mg/ml in the sperm cell composition.
5. The method of claim 4 , wherein the alpha-ketoglutarate is at a concentration ranging from 0.25 mg/ml to 0.45 mg/ml in the sperm cell composition.
6. The method of claim 1 , wherein the sperm cell composition further comprises vitamin B12, or a vitamin B12 vitamer.
7. The method of claim 4 , wherein the vitamin B12, or the vitamin B12 vitamer, is at a concentration ranging from 0.01 mg/ml to 1 mg/ml in the sperm cell composition.
8. The method of claim 1 , wherein the sperm cell composition further comprises a DNA selective dye.