System for in-vitro fertilization with spermatozoa separated into X-chromosome and Y-chromosome bearing populations

• US 7,094,527 B2
• Filed: 11/29/2001
• Issued: 08/22/2006
• Est. Priority Date: 11/29/2000
• Status: Expired due to Term

• Current Assignee: XY, LLC (Inguran LLC)
• Original Assignee: Colorado State University Research Foundation, XY Incorporated (Inguran LLC)
• Inventors: Seidel, George E., Lu, Kehuan, Suh, Tae Kwang
• Primary Examiner(s): TATE, CHRISTOPHER ROBIN
• Application Number:
  US10/433,191
• Publication Number:
  US 20040132001A1
• Time in Patent Office: 1,727 Days
• Field of Search: 435/2, 600/33, 600/34, 600/35
• US Class Current: 435/2
• CPC Class Codes: A01N 1/02 Preservation of living parts
• CPC Class Codes: A01N 1/0263 Non-refrigerated containers specially adapted for transporting or storing living parts whilst preserving, e.g. cool boxes, blood bags or "straws" for cryopreservation
• CPC Class Codes: A01N 1/0273 Transport containers A01N1/0268 takes precedence
• CPC Class Codes: A61D 19/00 Instruments or methods for reproduction or fertilisation
• CPC Class Codes: C12N 5/0604 Whole embryos; Culture medium therefor

Abstract

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).

Claims

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 μ

g 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.