Sperm cell processing systems
Semen and sperm cell processing and preservation systems, and methods of producing a mammal and methods of producing mammalian embryos are disclosed. The present invention is directed to sperm cell preservation, fertilization, and insemination, maintaining or enhancing sperm quality and addressing one or more sperm cell characteristics, such as viability, motility, functionality, fertilization rates, and pregnancy rates. Further, sperm cell characteristics may be addressed within the context of various collection, handling, separation, storage, transportation, usage, fertilization, or insemination techniques.
1. A method of producing a non-human mammalian embryo comprising:
obtaining sperm cells;
cryopreserving the sperm cells;
thawing the cryopreserved sperm cells;
staining the thawed sperm cells;
sorting the thawed sperm cells into X chromosome-bearing and/or Y chromosome bearing subpopulations by flow cytometry;
establishing at least one sperm sample from the sorted sperm cells;
cryopreserving the sorted sperm sample; and
establishing an increased blastocyst development rate using motile sperm cells from the cryopreserved sorted sperm sample compared to the blastocyst development rate using the obtained sperm cells.
2. The method of claim 1 , further comprising the step of producing a viable non-human mammal from the mammalian embryo.
3. The method of claim 1 , wherein the step of fertilizing at least one egg with the thawed, twice cryopreserved sperm cells through artificial insemination or in vitro fertilization further comprises fertilizing the at least one egg through artificial insemination.
4. The method of claim 3 , wherein the step of fertilizing the at least one egg through artificial insemination further comprises inseminating a superovulated female with the thawed, twice cryopreserved sperm cells.
5. The method of claim 1 , wherein the step of fertilizing at least one egg with the thawed, twice cryopreserved sperm cells through artificial insemination or in vitro fertilization further comprises fertilizing the at least one egg in vitro.
6. The method of claim 1 , further comprising the step of transferring the at least one fertilized egg to a recipient female.
7. The method of claim 1 , further comprising the step of obtaining at least one egg of a superovulated female of a species of the mammal and wherein the step of fertilizing includes fertilizing the at least one egg in vitro.
8. The method of claim 7 , wherein the step of fertilizing the at least one egg in vitro further comprises fertilizing more than one egg in vitro.
9. The method of claim 7 , further comprising the step of transferring the at least one fertilized egg to a recipient female.
10. The method of claim 1 , wherein the second cryopreserving step includes cryopreserving the at least one sperm sample at a concentration and dosage appropriate for artificial insemination or in vitro fertilization.
11. The method of claim 1 , wherein the step of establishing at least one sperm sample further comprises establishing a sample of the sorted, thawed sperm cells capable of fertilizing at least one egg of the female non-human mammal.
12. The method of claim 1 , wherein the step of establishing at least one sperm sample further comprises establishing a sample of the sorted, thawed sperm cells capable of fertilizing at least one egg of a superovulating female non-human mammal.
13. The method of claim of 1 , wherein the step of establishing at least one sperm sample further comprises establishing a sample of the sorted, thawed sperm cells capable of fertilization in vitro of at least one egg.
14. The method of claim 1 , further comprising the step of fertilizing a plurality of eggs of a female non-human mammal.
15. The method of claim 1 , further comprising the step of producing at least one animal of a pre-selected sex from the cryopreserved processed sperm cells.
16. The method of claim 1 , wherein the twice cryopreserved sperm cells have a pre-selected sex.
17. The method of claim 1 , wherein the twice cryopreserved sperm cells include a sperm sample selected from the group consisting of: a straw, a pellet, an insemination dose, and a prepared sperm sample.