System for in-vitro fertilization 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 comrprising a straw ( 19 ) and an incubation element ( 20 ) that can be sealed with a cap ( 22 ).
1. A method of transporting oocytes, comprising the steps of:
a. collecting oocytes from a female non-human mammal;
b. entraining the oocytes in a fertilization medium;
c. transferring the oocytes in said fertilization medium into at least one straw;
d. sealing said at least one straw containing said oocytes;
e. placing said at least one straw inside a closable incubation container capable of containing a separate, transportable incubation environment;
f. transferring said at least one straw inside of said incubation container;
g. establishing incubation conditions within said incubation container;
h. sealing said incubation container; and
i. transporting said oocytes from said female non-human mammal within said straw inside of said incubation container to another location.
2. A method of transporting oocytes as described in claim 1 , wherein said female non-human mammal is selected from the group consisting of non-human primates, bovids, ovids, equids, swine, and dolphins.
3. A method of transporting oocytes as described in claim 1 , wherein said fertilization medium comprises modified Tyrode's medium supplemented with 0.6 percent bovine serum albumin, 20 μg heparin per milliliter of Tyrode's medium, and a concentration of 5 milli-molar caffeine.
4. A method of transporting oocytes as described in claim 1 , wherein between about 10 and about 15 of said oocytes are contained within about 50 micro-liters of said fertilization medium.
5. A method of transporting oocytes as described in claim 1 , wherein said at least one straw has heat sealable aperture elements.
6. A method of transporting oocytes as described in claim 5 , wherein said at least one straw has a interior volume of about 0.25 milliliters.
7. A method of transporting oocytes as described in claim 1 , wherein said incubation container has sealable aperture elements.
8. A method of transporting oocytes as described in claim 7 , wherein said incubation container comprises a glass tube.
9. A method of transporting oocytes as described in claim 1 , wherein said incubation conditions comprise an atmosphere of 5 percent carbon dioxide in air and a temperature of 39 degrees Centigrade within said incubation container.
10. A method of transporting oocytes as described in claim 1 , further comprising the step of transferring sperm cells to said at least one straw containing said oocytes in said fertilization medium.
11. A method of transporting oocytes as described in claim 10 , wherein said step of transferring sperm cells to said at least one straw containing said oocytes comprises establishing a concentration of sperm cells in said fertilization medium of about 1 million to about 2 million per milliliter of fertilization medium.
12. A method of transporting oocytes as described in claim 1 , further comprising the step of separating spermatozoa into enriched X-chromosome bearing and Y-chromosome populations prior to said step of transferring the oocytes in said fertilization medium into said at least one straw.
13. A method of transporting oocytes as described in claim 12 , further comprising the step of transferring separated sperm cells to said at least one straw containing oocytes.
14. A method of transporting oocytes as described in claim 13 , wherein said step of transferring separated sperm cells to said at least one straw containing oocytes comprises establishing a concentration of said separated sperm cells in said fertilization medium of about 1 million to about 2 million per milliliter of fertilization medium.
15. A method of transporting oocytes as described in claims 10 , 11 , 12 , 13 , or 14 further comprising the step of transferring said at least one straw containing said oocytes to said incubation container.
16. A method of transporting oocytes as described in claim 15 , further comprising the step of establishing fertilization conditions within said incubation container.
17. A method of transporting oocytes as described in claim 16 , wherein said step of establishing fertilization conditions within said incubation container comprises 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.
18. A method of transporting oocytes as described in claim 16 , further comprising the step of transporting said oocytes in said fertilization conditions.
19. A method of transporting oocytes as described in claim 18 , further comprising the step of fertilizing at least some of said oocytes during transport.