In-vitro fertilization systems with spermatozoa separated into X-chromosome and Y-chromosome bearing populations
View Patent ↗An IVF system for successfully utilizing spermatozoa separated into X-chromosome bearing and into Y-chromosome bearing population for insemination. The IVF system includes fertilization medium that can shorten the time from insemination to cleavage and a portable incubator for the transportation of maturing oocytes and inseminated oocytes comprising a straw ( 19 ) and an incubation element ( 20 ) that can be sealed with a cap ( 22 ).
1. A method of in vitro fertilization of oocytes, comprising the steps of:
a) obtaining oocytes of a non-human mammal;
b) obtaining sperm cells of said non-human mammal separated into an X-chromosome bearing population or a Y-chromosome bearing population;
c) combining said oocytes of said non-human mammal in a fertilization medium supplemented with an amount of non-essential amino acids with said sperm cells of either said X-chromosome bearing population or said Y-chromosome bearing population;
d) establishing said plurality of oocytes with said sperm cells of said X-chromosome bearing population or said Y-chromosome bearing population in incubation conditions within an incubation element; and
e) fertilizing at least one of said oocytes of said non-human mammal with said sperm cells of said X-chromosome bearing population or said Y-chromosome bearing populations, said amount of non-essential amino acids sufficient to achieve an increased rate of development of the resulting at least one embryo.
2. The method of in vitro fertilization of oocytes of claim 1 , wherein said oocytes fertilized with said sperm cells of said X-chromosome bearing population or said Y-chromosome bearing population in said fertilization medium containing non-essential amino acids achieves an increased rate of development through at least the two cell stage.
3. The method of in vitro fertilization of oocytes of claim 2 , further comprising the step of entraining about 10 of said oocytes to about 15 of said oocytes in about 50 microliters of said fertilization medium.
4. The method of in vitro fertilization of oocytes of claim 3 , wherein said fertilization medium is selected from the group consisting of: a chemically defined medium, SOF, Tyrode's medium supplemented with Eagles Medium, Tyrode's medium supplemented with 0.6 percent bovine serum albumin, 20 μg heparin per milliliter, and a concentration of 5 milli-molar caffeine, and combinations thereof.
5. The method of in vitro fertilization of oocytes of claim 1 , wherein said incubation conditions comprise an atmosphere of 5 percent carbon dioxide in air at a temperature between about 37 degrees Centigrade and about 41 degrees Centigrade for a duration of about 18 hours to about 20 hours.
6. The method of in vitro fertilization of oocytes of claim 1 , further comprising the step of establishing a concentration of said sperm cells of said X-chromosome bearing population or said Y-chromosome bearing population in said fertilization medium of about 1 million to about 2 million per milliliter of said fertilization medium.
7. The method of in vitro fertilization of oocytes of claim 1 , further comprising the step of transferring said oocytes and said sperm cells into a straw.
8. The method of in vitro fertilization of oocytes of claim 7 , wherein said straw has heat sealable aperture elements.
9. The method of in vitro fertilization of oocytes of claim 8 , wherein said straw has an interior volume of about 0.25 milliliters.
10. The method of in vitro fertilization of oocytes of claim 7 , wherein said incubation element has sealable aperture elements.
11. The method of in vitro fertilization of oocytes of claim 10 , wherein said incubation element comprises a glass tube.
12. The method of in vitro fertilization of oocytes of claim 11 , wherein said non-human mammal is selected from the group consisting of primates, bovids, ovids, equids, swine, and dolphins.