In-vitro fertilization systems with spermatozoa separated into X-chromosome and Y-chromosome bearing populations

DC CAFC

US 8,569,053 B2
Filed: 02/24/2012
Issued: 10/29/2013
Est. Priority Date: 11/29/2000
Status: Expired due to Term

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First Claim

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1. A method of in-vitro fertilization of oocytes, comprising:

a) obtaining oocytes of a non-human mammal;

b) obtaining stained sperm cells of said non-human mammal, said stained sperm cells separated by a flow cytometer into an X-chromosome bearing population or a Y-chromosome bearing population;

c) combining said oocytes in a fertilization medium with said stained sperm cells separated by said flow cytometer, said stained sperm cells of either one of said X-chromosome bearing population or said Y-chromosome bearing population; and

d) fertilizing in-vitro at least one of said oocytes with said stained sperm cells of either one said X-chromosome bearing population or said Y-chromosome bearing population to produce sex-selected embryos capable of blastocyst formation.

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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 comprising a straw (19) and an incubation element (20) that can be sealed with a cap (22).

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18 Claims

1. A method of in-vitro fertilization of oocytes, comprising:
- a) obtaining oocytes of a non-human mammal;
  
  b) obtaining stained sperm cells of said non-human mammal, said stained sperm cells separated by a flow cytometer into an X-chromosome bearing population or a Y-chromosome bearing population;
  
  c) combining said oocytes in a fertilization medium with said stained sperm cells separated by said flow cytometer, said stained sperm cells of either one of said X-chromosome bearing population or said Y-chromosome bearing population; and
  
  d) fertilizing in-vitro at least one of said oocytes with said stained sperm cells of either one said X-chromosome bearing population or said Y-chromosome bearing population to produce sex-selected embryos capable of blastocyst formation.
- View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
- - 2. The method of in-vitro fertilization of oocytes of claim 1, wherein said fertilization medium contains an amount of non-essential amino acids.
  - 3. The method of in-vitro fertilization of oocytes of claim 2, further comprising sufficiently supplementing said fertilization medium with said non-essential amino acids to achieve an increased rate of development through at least a two cell stage.
  - 4. The method of in-vitro fertilization of oocytes of claim 3, wherein said non-human mammal is a bovine non-human mammal, and wherein said step of achieving said increased rate of development through at least said two cell stage is selected from the group consisting of:
    - achieving at least about a 20% increase in yield of said two cell stage;
      
      achieving at least about a 45% increase in yield of said two cell stage;
      
      achieving at least about a 70% increase in yield of said two cell stage;
      
      achieving at least about a 26% yield of said two cell stage;
      
      achieving at least about a 49% yield of said two cell stage; and
      
      achieving at least about a 75% yield of said two cell stage.
  - 5. The method of in-vitro fertilization of oocytes of claim 2, wherein said oocytes fertilized with said stained sperm cells of said X-chromosome bearing population or said Y-chromosome bearing population in said fertilization medium containing said non-essential amino acids achieve an increased rate of said sex-selected blastocysts suitable for embryo transfer.
  - 6. The method of in-vitro fertilization of oocytes of claim 5, wherein said non-human mammal is a bovine non-human mammal, and wherein said step of achieving said increased rate of said sex-selected blastocysts suitable for embryo transfer is selected from the group consisting of:
    - achieving at least about a 9% increase in said rate of blastocysts;
      
      achieving at least about a 12% increase in said rate of blastocysts;
      
      achieving at least about a 16% increase in said rate of blastocysts;
      
      achieving at least about a 52% yield of said blastocysts;
      
      achieving at least about a 61% yield of said blastocysts;
      
      achieving at least about a 64% yield of said blastocysts; and
      
      achieving at least about a 68% yield of said blastocysts.
  - 7. The method of in-vitro fertilization of oocytes of claim 5, wherein said non-human mammal comprises a bovine non-human mammal, and wherein said sex-selected blastocysts comprise Grade 1 blastocysts.
  - 8. The method of in-vitro fertilization of oocytes of claim 1, further comprising supplementing said fertilization medium with an amount of non-essential amino acids.
  - 9. The method of in-vitro fertilization of oocytes of claim 1, wherein said stained sperm cells comprise frozen-thawed stained sperm cells of said non-human mammal prior separated by said flow cytometer into said X-chromosome bearing population or said Y-chromosome hearing population.
  - 10. The method in-vitro fertilization of bovine oocytes as described in claim 1, further comprising establishing said plurality of oocytes with said stained sperm cells of said X-chromosome bearing population or said Y-chromosome bearing population in incubation conditions within an incubation element.
  - 11. The method of in-vitro fertilization of oocytes of claim 10, wherein said incubation conditions comprise an atmosphere of about 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.
  - 12. The method of in-vitro fertilization of oocytes of claim 11, further comprising transferring said oocytes and said stained sperm cells into a tubular member.
  - 13. The method of in-vitro fertilization of oocytes of claim 12, wherein said tubular member has heat sealable aperture elements.
  - 14. The method of in-vitro fertilization of oocytes of claim 13, wherein said tubular member has an interior volume of about 0.25 milliliters.
  - 15. The method of in-vitro fertilization of oocytes of claim 12, further comprising transferring said tubular member into an incubation element.
  - 16. The method of in-vitro fertilization of oocytes of claim 15, wherein said incubation element comprises a glass tube.
  - 17. The method of in-vitro fertilization of oocytes of claim 16, wherein said tubular member comprises an artificial insemination straw.
  - 18. The method of in-vitro fertilization of oocytes of claim 1, wherein said first and second non-human mammal is selected from the group consisting of non-human primates, bovids, ovids, equids, swine, and dolphins.

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Current Assignee
XY, LLC (Inguran LLC)
Original Assignee
XY, LLC (Inguran LLC)
Inventors
Seidel, George E., Lu, Kehuan, Suh, Tae Kwang
Primary Examiner(s)
DAVIS, RUTH A

Application Number

US13/404,706
Publication Number

US 20120164733A1
Time in Patent Office

613 Days
Field of Search

435/2, 435/325, 435/373, 435/363
US Class Current

435/325
CPC Class Codes

A01N 1/02   Preservation of living parts

A01N 1/0263   Non-refrigerated containers...

A01N 1/0273   Transport containers A01N1/...

A61D 19/00   Instruments or methods for ...

C12N 5/0604   Whole embryos; Culture medi...

In-vitro fertilization systems with spermatozoa separated into X-chromosome and Y-chromosome bearing populations

First Claim

8 Assignments

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Abstract

12 Citations

18 Claims

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In-vitro fertilization systems with spermatozoa separated into X-chromosome and Y-chromosome bearing populations

First Claim

8 Assignments

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Litigations

0 Petitions

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Accused Products

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Abstract

12 Citations

18 Claims

Specification

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Solutions

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